UDC 575.224.2
Íåõðîìîñîìíûé öèòîãåíåòè÷åñêèé àíàëèç
ñîìàòè÷åñêèõ êëåòîê ìëåêîïèòàþùèõ
Î. À. Êîâàëåâà, Í. À. Áåçäåíåæíûõ, Þ. È. Êóäðÿâåö
Èíñòèòóò ýêñïåðèìåíòàëüíîé ïàòîëîãèè, îíêîëîãèè è ðàäèîáèîëîãèè èì. Ð. Å. Êàâåöêîãî ÍÀÍ Óêðàèíû Óë. Âàñèëüêîâñêàÿ, 45, Êèåâ, Óêðàèíà, 03022
strukov2002@mail.ru
 îáçîðå ðàññìîòðåíû ìóòàöèîííûå ñîáûòèÿ, ïðîèñõîäÿùèå â ñîìàòè÷åñêèõ êëåòêàõ ìëåêîïèòàþùèõ ïîä âëèÿíèåì ðàçëè÷íûõ ýíäî- è ýêçîãåííûõ ôàêòîðîâ. Íåõðîìîñîìíûé ìåòîä èññëåäîâàíèÿ ïîçâîëÿåò ó÷èòûâàòü ñîâîêóïíîñòü õàðàêòåðèñòèê êëåòêè áåç òðóäîåìêîãî àíàëèçà íåïîñðåäñòâåííî ñàìèõ õðî-ìîñîì.  ðåçóëüòàòå ïîëó÷àþò èíôîðìàöèþ î ìèòîòè÷åñêîì (ôàçû ìèòîçà, êîëè÷åñòâî ÿäåð íà êëåò-êó, ìèêðîÿäðà, ïàòîëîãèè ìèòîçà) è âèòàëüíîì (ìèòîòè÷åñêèé èíäåêñ, àïîïòîç) ñòàòóñàõ êëåòêè, à òàêæå î ñîñòîÿíèè öåëîñòíîñòè õðîìîñîì (íàëè÷èå íóêëåîïëàçìàòè÷åñêèõ ìîñòîâ, ÿäåðíûõ ïðîòðó-çèé, ôðàãìåíòàöèè õðîìîñîì, ìèêðîÿäåð).  çàâèñèìîñòè îò èçó÷àåìîãî ìàòåðèàëà (ýðèòðîöèòû è ëèìôîöèòû ïåðèôåðè÷åñêîé êðîâè, êëåòêè áóêêàëüíîãî ýïèòåëèÿ, ïîñòîÿííûå êëåòî÷íûå ëèíèè è ò. ï.) ìîæíî âûáðàòü êîìïëåêñ öèòîãåíåòè÷åñêèõ õàðàêòåðèñòèê, íàèáîëåå èíôîðìàòèâíûé â êàæäîì îò-äåëüíîì ñëó÷àå äëÿ îïðåäåëåíèÿ ìóòàöèîííûõ ñïåêòðîâ â ñîìàòè÷åñêèõ êëåòêàõ ìëåêîïèòàþùèõ.
Êëþ÷åâûå ñëîâà: ìóòàãåíåç, ñîìàòè÷åñêàÿ êëåòêà, öèòîãåíåòè÷åñêèé àíàëèç, ïàòîëîãèÿ.
Èçó÷åíèå âëèÿíèÿ ôàêòîðîâ îêðóæàþùåé ñðåäû íà ãåíåòè÷åñêèé àïïàðàò ìëåêîïèòàþùèõ ÿâëÿåòñÿ îä-íîé èç îñíîâíûõ ïðîáëåì ýêîëîãè÷åñêîé ãåíåòèêè. Õðîìîñîìíûé àïïàðàò î÷åíü ÷óòêî ðåàãèðóåò íà ëþáîå âîçäåéñòâèå, íà÷èíàÿ îò ýëåêòðè÷åñêèõ ïðè-áîðîâ, âèðóñíîé èíôåêöèè è çàêàí÷èâàÿ çàãðÿçíå-íèåì îêðóæàþùåé ñðåäû. Ïîâûøåííàÿ öèòîãåíåòè-÷åñêàÿ íåñòàáèëüíîñòü ìîæåò áûòü ïðè÷èíîé áåñ-ïëîäèÿ è ðàçâèòèÿ îíêîëîãè÷åñêèõ çàáîëåâàíèé. Îöåíêà óðîâíÿ ñîìàòè÷åñêîãî ìóòàãåíåçà â íàñòîÿ-ùåå âðåìÿ ÿâëÿåòñÿ íàèáîëåå îáîñíîâàííûì è ïåð-ñïåêòèâíûì ïîäõîäîì ê ôîðìèðîâàíèþ ãðóïï âû-ñîêîãî êàíöåðîãåííîãî ðèñêà. Òðàäèöèîííî ìóòà-öèîííûé ïðîöåññ â ñîìàòè÷åñêèõ êëåòêàõ ÷åëîâåêà îöåíèâàþò ìåòîäîì ó÷åòà õðîìîñîìíûõ àáåððà-öèé, ÿâëÿþùèõñÿ îáùåïðèçíàííûì èíäèêàòîðîì ðàçíîãî ðîäà ìóòàãåííûõ âëèÿíèé, îäíàêî èõ ïîä-ñ÷åò òðåáóåò âûñîêîãî ïðîôåññèîíàëèçìà èññëåäî-âàòåëÿ è çàíèìàåò ìíîãî âðåìåíè.
Íåõðîìîñîìíûé ìåòîä èññëåäîâàíèÿ «öèòîìà» ïðåäïîëàãàåò âîçìîæíîñòü îïðåäåëåíèÿ ÷àñòîòû ìèêðîÿäåð, óðîâíÿ êëåòî÷íîé ãèáåëè è ìèòîòè÷åñ-êîé äèñôóíêöèè [1, 2]. Ïîä «öèòîìîì» ïîäðàçóìå-âàåòñÿ ñîâîêóïíîñòü õàðàêòåðèñòèê êëåòêè, âêëþ-÷àþùàÿ åå âèòàëüíûé (íåêðîç, àïîïòîç) è ìèòîòè-÷åñêèé (ìèêðîÿäðà, ìåòàôàçû, àíàôàçû, îäíî- è äâó-ÿäåðíîñòü) ñòàòóñû, à òàêæå ñîñòîÿíèå õðîìîñîì-íîé öåëîñòíîñòè (íàëè÷èå ìèêðîÿäåð, íóêëåîñîì-íûõ ìîñòîâ, ÿäåðíóêëåîñîì-íûõ ïî÷åê, ðàñïðåäåëåíèÿ õðîìî-ñîìîñïåöèôè÷íûõ ñèãíàëîâ).  äàííîì îáçîðå ïðåä-ñòàâëåíû âñåâîçìîæíûå âàðèàíòû öèòîãåíåòè÷åñ-êèõ íàðóøåíèé, êîòîðûå ìîæíî ó÷èòûâàòü ïðè îöåí-êå ãåíåòè÷åñêîãî áëàãîïîëó÷èÿ ó ëèö, ïîäâåðãøèõ-ñÿ âëèÿíèþ íåáëàãîïðèÿòíûõ ôàêòîðîâ îêðóæàþ-ùåé ñðåäû, â òîì ÷èñëå íà ïðîèçâîäñòâå. Åñòåñòâåí-íî, â çàâèñèìîñòè îò èçó÷àåìîé òêàíè (ýðèòðîöèòû è ëèìôîöèòû ïåðèôåðè÷åñêîé êðîâè, êëåòêè áóê-êàëüíîãî ýïèòåëèÿ è ò. ä.) èññëåäîâàòåëü ìîæåò âû-áðàòü öèòîãåíåòè÷åñêèå õàðàêòåðèñòèêè, íàèáîëåå èíôîðìàòèâíûå äëÿ êàæäîãî îòäåëüíîãî ñëó÷àÿ.
ISSN 0233–7657. Biopolymers and Cell. 2013. Vol. 29. N 1. P. 33–41 doi: 10.7124/bc.000803
Äàííàÿ èíôîðìàöèÿ ìîæåò áûòü ïîëåçíà è äëÿ öèòîãåíåòèêîâ, ðàáîòàþùèõ ñ ïîñòîÿííûìè êëå-òî÷íûìè êóëüòóðàìè.
Ïàòîëîãèè ìèòîçà. Â ëèòåðàòóðå îïèñàíû òðè
êðèòåðèÿ äëÿ âûÿâëåíèÿ íàðóøåíèé ìèòîòè÷åñêîé àêòèâíîñòè â ñîìå: 1) íåïðåêðàùàþùååñÿ äåëåíèå; 2) ìíîãîïîëþñíîñòü àíàôàçû è 3) èçìåíåíèå ñîäåð-æàíèÿ ÄÍÊ. Êàæäîå ñîáûòèå ìîæíî ïðîñëåæèâàòü, èñïîëüçóÿ ìåòîäû ñâåòîâîé ìèêðîñêîïèè [3]. Ïðî-öåññ ìèòîòè÷åñêîãî äåëåíèÿ êëåòîê î÷åíü ÷óâñòâè-òåëåí ê äåéñòâèþ ñàìûõ ðàçíîîáðàçíûõ ôàêòîðîâ. Âîçíèêàþùèå ïàòîëîãè÷åñêèå ìèòîçû âêëþ÷àþò â ñåáÿ ôðàãìåíòàöèþ õðîìîñîì, õðîìàòèäíûå ìîñòû, îòñòàâàíèå õðîìîñîì â ìåòàêèíåçå, îòñòàâàíèå õðî-ìîñîì ïðè ðàñõîæäåíèè ê ïîëþñàì, ðàññåèâàíèå ãèïåðñïèðàëèçîâàííûõ õðîìîñîì, ìíîãîïîëþñíûå ìèòîçû, ïîëûå ìåòàôàçû.
Ôðàãìåíòàöèÿ è ïóëüâåðèçàöèÿ õðîìîñîì ÿâëÿ-þòñÿ îñíîâíîé ôîðìîé ìèòîòè÷åñêîé ñìåðòè êëåò-êè, êîòîðàÿ èäåíòèôèöèðóåòñÿ îáùèì öèòîãåíåòè-÷åñêèì àíàëèçîì (ðèñ. 1, ñì. âêëåéêó – âñå êëåòî÷-íûå ëèíèè, èñïîëüçîâàíêëåòî÷-íûå â ýêñïåðèìåíòàõ, ïîëó-÷åíû èç Êëåòî÷íîãî áàíêà ëèíèé èç òêàíåé ÷åëîâå-êà è æèâîòíûõ Èíñòèòóòà ýêñïåðèìåíòàëüíîé ïà-òîëîãèè, îíêîëîãèè è ðàäèîáèîëîãèè èì. Ð. Å. Êà-âåöêîãî ÍÀÍ Óêðàèíû). Îá îòëè÷èè ýòîé ïàòîëî-ãèè îò àïîïòîçà ñâèäåòåëüñòâóþò ðàçëè÷íûå èññëå-äîâàíèÿ, â òîì ÷èñëå è áèîõèìè÷åñêèå [4].
Ìîðôîëîãè÷åñêè ýòó ïàòîëîãèþ ìîæíî ðàçäå-ëèòü, ïî êðàéíåé ìåðå, íà òðè ãðóïïû: ðàííÿÿ ôðàã-ìåíòàöèÿ õðîìîñîì, ïðè êîòîðîé ðàçðóøåíû íå-ñêîëüêî õðîìîñîì; ñðåäíÿÿ ñòàäèÿ äðîáëåíèÿ, íà êîòîðîé ôðàãìåíòèðîâàíî çíà÷èòåëüíîå ÷èñëî õðî-ìîñîì; ïîçäíÿÿ ñòàäèÿ äðîáëåíèÿ, êîãäà âñå èëè áîëüøèíñòâî õðîìîñîì ôðàãìåíòèðîâàíû. Èñõîäÿ èç ýòîãî ìîæíî ïðåäïîëîæèòü, ÷òî õðîìîñîìíàÿ ôðàãìåíòàöèÿ – ýòî ïðîãðåññèâíûé ïðîöåññ ïîñòå-ïåííîé äåãðàäàöèè õðîìîñîì [4]. Ïðè ïóëüâåðèçà-öèè õðîìîñîìû ðàñïàäàþòñÿ íà î÷åíü ìåëêèå ôðàã-ìåíòû, ïðè÷åì áîëüøèíñòâî èç íèõ ëèøåíû êèíå-òîõîðà è ïîýòîìó îñòàþòñÿ íåïîäâèæíûìè. Ñïîñîá-íîñòüþ ê èíäóêöèè ïóëüâåðèçàöèè õðîìîñîì îáëà-äàþò âèðóñû êîðè, ïàðîòèòà, èñòèííîé ÷óìû ïòèö, áîëåçíè Íüþêàñëà, àäåíîâèðóñ 12-ãî òèïà, ïðîñòî-ãî ãåðïåñà è êëåùåâîïðîñòî-ãî ýíöåôàëèòà [5]. Èíêóáàöèÿ êóëüòóðû êëåòîê HeLa è Lu-106 ñ Í3-òèìèäèíîì
ïîçâîëèëà óñòàíîâèòü, ÷òî ïóëüâåðèçàöèÿ âîçíèêà-åò íà S-ñòàäèè êëâîçíèêà-åòî÷íîãî öèêëà. Õðîìîñîìû, íà ìî-ìåíò âîçäåéñòâèÿ âèðóñà ïðîøåäøèå ïåðèîä ðå-ïëèêàöèè, íå ïîäâåðãàþòñÿ ïóëüâåðèçàöèè [6].  ëè-òåðàòóðå èìåþòñÿ äàííûå, ïîçâîëÿþùèå ñ÷èòàòü, ÷òî ïóëüâåðèçàöèÿ ÿâëÿåòñÿ ñëåäñòâèåì ëîêàëüíîé êîíäåíñàöèè õðîìîñîì. Ñòåíìàí è Ñàêñåëà [7] ïî-ëàãàþò, ÷òî ïóëüâåðèçàöèþ ïðàâèëüíåå áûëî áû íà-çûâàòü ïðåæäåâðåìåííîé êîíäåíñàöèåé õðîìîñîì.
 äðóãèõ ñëó÷àÿõ ïàòîëîãèè ìèòîçà â ðåçóëüòà-òå îáúåäèíåíèÿ äâóõ ïîâðåæäåííûõ õðîìîñîì ôîð-ìèðóåòñÿ äèöåíòðè÷åñêàÿ õðîìîñîìà. Ïîä âîçäåé-ñòâèåì îáîèõ ìèòîòè÷åñêèõ öåíòðîâ õðîìîñîìà ðàñ-òÿãèâàåòñÿ ìåæäó íèìè è îáðàçóåò ìîñò, ÷òî âëèÿåò íà òå÷åíèå çàêëþ÷èòåëüíûõ ñòàäèé ìèòîçà è çàäåð-æèâàåò öèòîòîìèþ (ðèñ. 2, ñì. âêëåéêó).
Îòñòàâàíèå õðîìîñîì â ìåòàêèíåçå è ïðè ðàñ-õîæäåíèè ê ïîëþñàì âîçíèêàåò ïðè ïîâðåæäåíèè êèíåòîõîðà è/èëè âåðåòåíà äåëåíèÿ (ðèñ. 3, ñì. âêëåéêó).
Íàèáîëåå ÷àñòî âñòðå÷àåòñÿ îñòàíîâêà ìèòîçà íà ñòàäèè ìåòàôàçû. Ýòî ïðîèñõîäèò â ðåçóëüòàòå èçìåíåíèé âåðåòåíà äåëåíèÿ. Ìíîãèå âåùåñòâà, áëî-êèðóþùèå ìèòîç, ñðåäè êîòîðûõ òàêèå öèòîñòàòè-êè, êàê êîëõèöèí è êîëöåìèä, ïðåïÿòñòâóþò ïîëè-ìåðèçàöèè òóáóëèíîâ. Âñëåäñòâèå ýòîãî íîâûå ìèê-ðîòðóáî÷êè âåðåòåíà íå îáðàçóþòñÿ, à ãîòîâûå ïîë-íîñòüþ ðàçáèðàþòñÿ. Ïðè ýòîì ìèòîòè÷åñêèå õðî-ìîñîìû ñîáèðàþòñÿ â öåíòðå êëåòêè, íî íå ôîðìè-ðóþò ìåòàôàçíîé ïëàñòèíêè, à ðàñïîëàãàþòñÿ áåç âñÿêîãî ïîðÿäêà (Ê-ìèòîç).
Ê ñõîäíûì âûâîäàì ïðèâîäèò äåéñòâèå íà êëåò-êó èíãèáèòîðîâ ñèíòåçà ÀÒÔ (íèòðîôåíîë, îëèãî-ìèöèí) è ðÿäà ÿäîâèòûõ âåùåñòâ (ìåðêàïòîýòàíîë). Åñëè äåéñòâèå ýòèõ ôàêòîðîâ êðàòêîâðåìåííî, òî âîçìîæíû âîññòàíîâëåíèå ìèêðîòðóáî÷åê âåðåòåíà è êëåòî÷íîå äåëåíèå. Ïðè óìåðåííûõ âîçäåéñòâèÿõ êëåòêè ìîãóò íå ïîãèáíóòü, à áåç ìèòîçà âñòóïàòü â ñëåäóþùèé êëåòî÷íûé öèêë.  ýòîì ñëó÷àå íåðàçî-øåäøèåñÿ õðîìîñîìû äåêîíäåíñèðóþòñÿ, îáðàçó-þòñÿ íîâàÿ ÿäåðíàÿ îáîëî÷êà è íîâîå, íî óæå òåòðà-ïëîèäíîå ÿäðî.
Òàê âîçíèêàþò ïîëèïëîèäíûå êëåòêè ïîä âëèÿ-íèåì êîëõèöèíà. Âîññòàíîâëåíèå íîðìàëüíîãî ìè-òîçà ïîñëå åãî áëîêèðîâàíèÿ êîëõèöèíîì ñâÿçàíî ñ äîïîëíèòåëüíûìè ñòàäèÿìè ñèíòåçà áåëêà
ìèðîâàíèå ìèêðîòðóáî÷åê âåðåòåíà äåëåíèÿ). Âåðî-ÿòíî, àêòèâíîñòü ïðîöåññîâ ðåôîðìèðîâàíèÿ è ñáîð-êà ìèêðîòðóáî÷åê âî âðåìÿ âîññòàíîâëåíèÿ çàâèñÿò îò ñòåïåíè èõ ðàçðóøåíèÿ è õàðàêòåðà ïîâðåæäàþ-ùåãî àãåíòà [8]. Ê îäíîìó èç âèäîâ Ê-ìèòîçà îòíî-ñÿò áåñïîðÿäî÷íîå ðàññåèâàíèå ãèïåðñïèðàëèçî-âàííûõ õðîìîñîì â ìåòàôàçå.
Ê àíîìàëèÿì äåëåíèÿ êëåòîê ïðèíàäëåæàò è ìíîãîïîëþñíûå ìèòîçû.  ýòîì ñëó÷àå â ìåòàôàçå îáðàçóåòñÿ íå áèïîëÿðíîå âåðåòåíî, à âåðåòåíî ñ òðå-ìÿ èëè ÷åòûðüòðå-ìÿ ïîëþñàìè. Òàêàÿ àíîìàëèÿ îáó-ñëîâëåíà íàðóøåíèÿìè ôóíêöèé öåíòðèîëåé: äè-ïëîñîìà ðàñïàäàåòñÿ íà äâå àêòèâíûå ìîíîöåíòðèî-ëè. Ýòè èçìåíåíèÿ ìîãóò ïðîèñõîäèòü ñïîíòàííî (÷òî õàðàêòåðíî äëÿ îïóõîëåâûõ êëåòîê) èëè ïîñëå âîçäåéñòâèÿ ðàçëè÷íûõ èíãèáèòîðîâ ìèòîòè÷åñêîãî äåëåíèÿ. Óêàçàííûå àíîìàëüíûå ìíîãîïîëþñíûå ìèòîòè÷åñêèå ôèãóðû (ðèñ. 4, ñì. âêëåéêó) ñïîñîá-íû âñòóïàòü â àíàôàçó è ó÷àñòâîâàòü â ðàñõîæäåíèè õðîìîñîì ê ïîëþñàì, âñëåä çà ÷åì ìîæåò íàñòó-ïèòü öèòîòîìèÿ ñ îáðàçîâàíèåì òðåõ, ÷åòûðåõ è áî-ëåå êëåòîê.  ýòèõ ñëó÷àÿõ ðàâíîìåðíîãî ðàñïðåäå-ëåíèÿ õðîìîñîì íå ïðîèñõîäèò, à îáðàçîâàâøèåñÿ êëåòêè ñîäåðæàò ñëó÷àéíûå è óìåíüøåííûå íàáî-ðû õðîìîñîì. Ñôîðìèðîâàâøèåñÿ àíåóïëîèäíûå êëåòêè îáû÷íî áûñòðî ïîãèáàþò.
Ôèãóðû, ïðåäñòàâëåííûå íà ðèñ. 4, ñâèäåòåëü-ñòâóþò íå òîëüêî î «ðàñùåïëåíèè âåðåòåíà» (öåíò-ðèîëåé), íî è î íàðóøåíèÿõ îòäåëüíûõ íèòåé âåðå-òåíà èëè öåíòðîìåð õðîìîñîì, â ðåçóëüòàòå ÷åãî îòäåëüíûå õðîìîñîìû íå âõîäÿò â îáðàçîâàâøèåñÿ ãðóïïû.
Ìíîãîïîëþñíûé ìèòîç ìîæíî ðàññìàòðèâàòü äâîÿêî: êàê ñïîñîá ñíèæåíèÿ ïëîèäíîñòè êëåòî÷-íîé ïîïóëÿöèè â íîðìå (ïðè êîíòðîëèðóåìîì ðàñ-ïðåäåëåíèè õðîìîñîìíûõ íàáîðîâ) [9] èëè êàê îä-íó èç ïðè÷èí íàðàñòàíèÿ àíåóïëîèäèè, íàïðèìåð, îïóõîëåâûõ êëåòîê (ïðè îòñóòñòâèè êîíòðîëÿ ðàñ-ïðåäåëåíèÿ õðîìîñîìíûõ íàáîðîâ) [10]. Ïîýòîìó èññëåäîâàíèå çàêîíîìåðíîñòåé îáðàçîâàíèÿ ìíîãî-ïîëþñíîãî ìèòîòè÷åñêîãî àïïàðàòà èìååò êàê îá-ùåáèîëîãè÷åñêîå, òàê è ïðàêòè÷åñêîå çíà÷åíèå.
Àíîìàëèè ìèòîòè÷åñêîãî äåëåíèÿ ìîãóò îïðå-äåëÿòüñÿ íàðóøåíèÿìè öèòîòîìèè.  ýòîì ñëó÷àå âîçíèêàþò äâóÿäåðíûå è ìíîãîÿäåðíûå êëåòêè, ÷òî ñâÿçàíî ñ ïîäàâëåíèåì îáðàçîâàíèÿ àêòèíîâûõ
ìèê-ðîôèëàìåíòîâ, ó÷àñòâóþùèõ â îáðàçîâàíèè êëåòî÷-íîé ïåðåòÿæêè â êîíöå òåëîôàçû (ðèñ. 5, ñì. âêëåéêó). Ïîëàÿ ìåòàôàçà èìååò âèä êîëüöà õðîìîñîì, êî-òîðûå ñîáèðàþòñÿ â ìåòàôàçíóþ ïëàñòèíêó, ðàñïî-ëàãàÿñü ïî ïåðèôåðèè êëåòêè (ðèñ. 6, ñì. âêëåéêó).
Íåðåäêî âñòðå÷àþòñÿ êîìáèíèðîâàííûå ôîðìû ïàòîëîãèè ìèòîçà (íàïðèìåð, ìíîãîïîëþñíûå ìè-òîçû ñ îòñòàâàíèåì õðîìîñîì èëè ñ ìîñòàìè è ò. ä.) (ðèñ. 7, ñì. âêëåéêó).
Âñëåäñòâèå ïàòîëîãè÷åñêèõ ìèòîçîâ âîçíèêàþò ìóòàöèè, ïîÿâëÿþòñÿ õðîìîñîìíûå àáåððàöèè, èã-ðàþùèå âåñüìà ñóùåñòâåííóþ ðîëü â íåîïëàñòè-÷åñêîé òðàíñôîðìàöèè êëåòîê. Ïàòîëîãè÷åñêèå ìèòîçû â íîðìàëüíûõ òêàíÿõ âñòðå÷àþòñÿ â 2–3 % êëåòîê, â òî âðåìÿ êàê â îïóõîëåâûõ – â 30–46 % êëåòîê. Óñòàíîâëåíî, ÷òî ÷åì áîëåå âûðàæåíà ìîð-ôîëîãè÷åñêàÿ àíàïëàçèÿ îïóõîëåâûõ êëåòîê è ÷åì âûøå ìèòîòè÷åñêàÿ àêòèâíîñòü êëåòîê, òåì ÷àùå âñòðå÷àþòñÿ ïàòîëîãè÷åñêèå ìèòîçû.
Ìíîãîÿäåðíûå êëåòêè è àìèòîç.
Ìíîãîÿäåð-íûå êëåòêè ðåãèñòðèðóþò ïðè ïàòîëîãè÷åñêèõ ïðî-öåññàõ, îáóñëîâëèâàþùèõ ðàçâèòèå ìíîãèõ çàáîëå-âàíèé [11, 12]. Ðàçðàáîòêà ïåðñïåêòèâíûõ ìåòîäîâ äèàãíîñòèêè è êîððåêöèè çàáîëåâàíèé çàâèñèò îò ïîíèìàíèÿ ñóòè ìåõàíèçìîâ îáðàçîâàíèÿ ìíîãî-ÿäåðíûõ êëåòîê è îò èõ ñâÿçè ñ ïðè÷èíàìè, ëåæà-ùèìè â îñíîâå ïàòîëîãè÷åñêîãî ïðîöåññà. Ôîðìè-ðîâàíèå ïîëèíóêëåàðîâ ïðîèñõîäèò çà ñ÷åò ðåàëè-çàöèè ÷åòûðåõ ìåõàíèçìîâ: â ðåçóëüòàòå ñëèÿíèÿ îäíîÿäåðíûõ êëåòîê [13], â ñëó÷àå áëîêàäû öèòîêè-íåçà [14], âñëåäñòâèå ìíîãîïîëþñíûõ ìèòîçîâ [15] è ïðè àìèòîòè÷åñêîì äåëåíèè ÿäðà [16].
 îòëè÷èå îò ïåðâûõ òðåõ õîðîøî èçó÷åííûõ ìå-õàíèçìîâ àìèòîç äîñòàòî÷íî ðåäêî âûñòóïàåò â êà-÷åñòâå îáúåêòà èññëåäîâàíèÿ è îáúåì èíôîðìà-öèè ïî ýòîìó âîïðîñó êðàéíå îãðàíè÷åí. Àìèòîç îò-ìå÷åí â êëåòêàõ ðàçëè÷íîãî ïðîèñõîæäåíèÿ [17– 19], â èíòàêòíûõ êëåòêàõ, êóëüòèâèðóåìûõin vitro [18], è â êëåòêàõ îïóõîëåé [19]. Îí èìååò âàæíîå çíà÷åíèå äëÿ ôîðìèðîâàíèÿ ìíîãîÿäåðíûõ êëåòîê [16] è ïðåäñòàâëÿåò ñîáîé ñòàäèéíûé ïðîöåññ [20], â õîäå êîòîðîãî ïîñëåäîâàòåëüíî ïðîèñõîäÿò ðàñ-òÿæåíèå ÿäðà, èíâàãèíàöèÿ êàðèîëåììû è ïåðåòÿæ-êà ÿäðà íà ÷àñòè [21] (ðèñ. 8, ñì. âêëåéêó).
Õîòÿ îáúåì äîñòîâåðíîé èíôîðìàöèè î ìîëå-êóëÿðíûõ è ñóáêëåòî÷íûõ ìåõàíèçìàõ àìèòîçà
Figures to article by O. A. Kovalova et al.
à á â ã
Ðèñ. 1. Ðàçíûå âàðèàíòû ôðàãìåíòàöèè è ïóëüâåðèçàöèè õðîìîñîì:à,á– ñðåäíÿÿ ñòàäèÿ äðîáëåíèÿ (ôðàãìåíòàöèè);â,ã– ïîçäíÿÿ ñòà-äèÿ äðîáëåíèÿ ñ ìåëêèìè ôðàãìåíòàìè – ïóëüâåðèçàöèÿ. Êëåòî÷íàÿ ëèíèÿ Ê562 (õðîíè÷åñêàÿ ìèåëîèäíàÿ ëåéêåìèÿ ÷åëîâåêà).´1000
à á â
Ðèñ. 3. Îòñòàâàíèå õðîìîñîì â ìåòàôàçå (à– êëåòî÷íàÿ ëèíèÿ À-549;á– êëåòî÷íàÿ ëèíèÿ Du-145 – êàðöèíîìà ïðîñòàòû ÷åëîâåêà) è â àíàôàçå (â– êëåòî÷íàÿ ëèíèÿ ÌÊÑ – ðàê ìîëî÷íîé æåëåçû êðû-ñû).´1000
à á â
Ðèñ. 4. Ìíîãîïîëþñíûå ìè-òîòè÷åñêèå ôèãóðû:à– òðåõ-ãðóïïîâàÿ ôèãóðà;á,â– ìíî-ãîãðóïïîâûå ôèãóðû. Êëå-òî÷íàÿ ëèíèÿ CMS-180, ïî-ëó÷åííàÿ èç îïóõîëåâîãî øòàììà S-180 – ñïîíòàííàÿ êàðöèíîìà ìûøè. ´1000
à á â
Ðèñ. 5. Äâóÿäåðíûå êëåòêè: à,â– áåç ìèêðîÿäðà;á– ñ ìèêðîÿäðîì. Êëåòî÷íàÿ ëè-íèÿ À-549.´1000
Ðèñ. 2. Õðîìàòèäíûé ìîñò. Êëå-òî÷íàÿ ëèíèÿ À-549 (êàðöèíîìà ëåãêîãî ÷åëîâåêà).´1000
Ðèñ. 7. Ìíîãîïîëþñíûé ìè-òîç ñ õðîìàòèäíûì ìîñòîì. Êëåòî÷íàÿ ëèíèÿ ÌÊÑ. ´1000
äîñòàòî÷åí, âñå æå èìåþòñÿ ñâåäåíèÿ îá ó÷àñòèè êëåòî÷íîãî öåíòðà â ðåàëèçàöèè äàííîãî ïðîöåññà. Òàêæå èçâåñòíî, ÷òî åñëè ÿäðà ñåãìåíòèðóþòñÿ ïîä äåéñòâèåì ìèêðîôèëàìåíòîâ è ìèêðîòðóáî÷åê, òî ðîëü ýëåìåíòîâ öèòîñêåëåòà íå èñêëþ÷åíà è â àìè-òîòè÷åñêîì äåëåíèè.
 ëèòåðàòóðå åñòü òàêæå äàííûå î òîì, ÷òî íå-ìèòîòè÷åñêîå äåëåíèå ÿäðà – ýòî ïðîöåññ, áëàãîäà-ðÿ êîòîðîìó êëåòêè èçáàâëÿþòñÿ îò èçáûòî÷íî àìï-ëèôèöèðîâàííîé ÄÍÊ è ÷òî òàêèå äåôåêòû êëåòîê ïðèâîäÿò ê ôîðìèðîâàíèþ ìèêðîÿäåð [22] (ðèñ. 9, ñì. âêëåéêó).
Î ôóíäàìåíòàëüíîì çíà÷åíèè àìèòîçà â ðåàëè-çàöèè âíóòðèêëåòî÷íûõ ïðîöåññîâ ñâèäåòåëüñòâó-åò ôàêò åãî ñóùåñòâîâàíèÿ âî ìíîãèõ òèïàõ êëñâèäåòåëüñòâó-åòîê è ïðè ðàçíûõ óñëîâèÿõ. Òàê êàê ðîëü àìèòîòè÷åñêî-ãî äåëåíèÿ ïîëèïëîèäíûõ ÿäåð â îáðàçîâàíèè ïî-ëèíóêëåàðîâ ñ÷èòàåòñÿ äîêàçàííîé, òî îñíîâíîé ñìûñë àìèòîçà ñîñòîèò â óñòàíîâëåíèè îïòèìàëü-íûõ ÿäåðíî-öèòîïëàçìàòè÷åñêèõ îòíîøåíèé, ïî-çâîëÿþùèõ êëåòêàì àäåêâàòíî îñóùåñòâëÿòü ðàç-íîîáðàçíûå ôóíêöèè [16].
Ìèêðîÿäåðíûé òåñò.  íà÷àëå 70-õ ãã. ÕÕ âåêà
Õåääë è Øìèä íåçàâèñèìî äðóã îò äðóãà ïðåäëî-æèëè ìèêðîÿäåðíûé òåñò, îñíîâàííûé íà ó÷åòå ìèê-ðîÿäåð â ïîëèõðîìàòîôèëüíûõ ýðèòðîöèòàõ êîñò-íîãî ìîçãà [23, 24]. Ïåðâîíà÷àëüíî îí áûë ðàçðàáî-òàí äëÿ ýðèòðîèäíûõ êëåòîê êîñòíîãî ìîçãà, à ïîç-æå åãî ñòàëè ïðèìåíÿòü äëÿ ó÷åòà ìèêðîÿäåð â ðàí-íèõ ñïåðìàòèäàõ, ïå÷åíè ïëîäà ïðè èçó÷åíèè òðàíñ-ïëàöåíòàðíîé àêòèâíîñòè õèìè÷åñêèõ ñîåäèíåíèé, â êëåòêàõ ñëèçèñòîé ðòà, ëèìôîöèòàõ ÷åëîâåêà, â êëåòêàõ ïå÷åíè è òîëñòîé êèøêè æèâîòíûõ. Ê íà-ñòîÿùåìó âðåìåíè ó÷åò ìèêðîÿäåð ñòàë âîçìîæåí â áîëüøèíñòâå ïîïóëÿöèé äåëÿùèõñÿ êëåòîê. Ïîêà-çàíî, ÷òî ìèêðîÿäåðíûé òåñò ïî ÷óâñòâèòåëüíîñòè íå óñòóïàåò òåñòó ïî âûÿâëåíèþ õðîìîñîìíûõ àáåð-ðàöèé â êëåòêàõ êîñòíîãî ìîçãà æèâîòíûõ, ÿâëÿÿñü îäíîâðåìåííî ãîðàçäî ìåíåå òðóäîåìêèì.
Ìèêðîÿäðà îáðàçóþòñÿ â ïðîöåññå êëåòî÷íûõ äåëåíèé èç îòñòàþùèõ àöåíòðè÷åñêèõ ôðàãìåíòîâ, âîçíèêøèõ ïðè ðàçðûâå õðîìîñîì (êëàñòîãåííûé ýôôåêò), è îòñòàþùèõ õðîìîñîì (àíåóãåííûé ýô-ôåêò). Ìèêðîÿäðà ìîãóò òàêæå ôîðìèðîâàòüñÿ çà ñ÷åò íåìèòîòè÷åñêîé ýêñòðóçèè õðîìàòèíà èç èí-òåðôàçíîãî ÿäðà [22]. Îíè íå ñîäåðæàò ÿäåðíîé
îáî-ëî÷êè, ñïîñîáíû ê ëèçèñó èëè ìîãóò âêëþ÷àòüñÿ â ÿäðî ïðè ïîñëåäóþùåì ìèòîçå [5] (ðèñ. 10, ñì. âêëåéêó).
Ìèêðîÿäåðíûé òåñò ÿâëÿåòñÿ îáùåïðèíÿòûì öèòîãåíåòè÷åñêèì ñïîñîáîì îöåíêè ìóòàãåííîãî äåéñòâèÿ àãåíòîâ ðàçëè÷íîé ïðèðîäû. Ñ åãî ïî-ìîùüþ íà ìóòàãåííóþ àêòèâíîñòü ïðîâåðåíî áîëü-øîå êîëè÷åñòâî õèìè÷åñêèõ, ôèçè÷åñêèõ è áèîëî-ãè÷åñêèõ àãåíòîâ [25–27], òåñò ïðèìåíÿþò óæå íà ïåðâîì ýòàïå âûÿâëåíèÿ ïîòåíöèàëüíûõ ìóòàãåíîâ è êàíöåðîãåíîâ. Ê åãî ïðåèìóùåñòâàì ñëåäóåò îò-íåñòè áûñòðîòó èññëåäîâàíèÿ íåçàâèñèìî îò êàðèî-òèïà âèäà, íåðåäêî õàðàêòåðèçóþùåãîñÿ çíà÷èòåëü-íûì êîëè÷åñòâîì ìåëêèõ è ïëîõî ðàçëè÷èìûõ õðî-ìîñîì, íàäåæíîñòü, à òàêæå âîçìîæíîñòü ïðîâåäå-íèÿ òåñòèðîâàïðîâåäå-íèÿ â òêàíÿõ ñ íèçêîé ìèòîòè÷åñêîé àêòèâíîñòüþ. Ìèêðîÿäåðíûé àíàëèç ïðîâîäÿò â êëåòêàõ ýìáðèîíîâ, â ñïåðìàòèäàõ, îîòèäàõ, ÷òî îñî-áåííî âàæíî ïðè ïðîãíîçå ïîñëåäñòâèé äëÿ áóäó-ùåãî ïîòîìñòâà.
Èñïîëüçîâàíèå ìèêðîÿäåðíîãî òåñòà â ñëóùè-âàþùèõñÿ êëåòêàõ ÷åëîâåêà – ñàìûé ÷óâñòâèòåëü-íûé è áûñòðûé ìåòîä âûÿâëåíèÿ äåéñòâèÿ ìóòàãå-íà/êàðöèíîãåíà, òàê æå êàê è àíòèìóòàãåíà/àíòè-êàðöèíîãåíà íà ÷åëîâå÷åñêèé îðãàíèçìin vivo[28]. Èññëåäîâàíèå êëåòîê áóêêàëüíîãî ýïèòåëèÿ – íàè-áîëåå óäîáíûé ñïîñîá ó÷åòà öèòîãåíåòè÷åñêèõ àíî-ìàëèé äëÿ ëþäåé, ïðîæèâàþùèõ íà íåáëàãîïðèÿò-íûõ òåððèòîðèÿõ èëè ðàáîòàþùèõ íà âðåäíåáëàãîïðèÿò-íûõ ïðî-èçâîäñòâàõ [29].
Àíàëèç ìèêðîÿäåð â ïîëèõðîìàòîôèëüíûõ ýðèò-ðîöèòàõ – ìåíåå ðàñïðîñòðàíåííûé òåñò, îäíàêî íå ìåíåå èíôîðìàòèâíûé [26]. Íàïðèìåð, óâåëè÷åíèå êîëè÷åñòâà ýðèòðîöèòîâ ñ ìèêðîÿäðàìè â ïåðèôå-ðè÷åñêîé êðîâè ó áîëüíûõ àòîïè÷åñêîé áðîíõèàëü-íîé àñòìîé ïðÿìî ïðîïîðöèîíàëüíî ñòåïåíè òÿæå-ñòè çàáîëåâàíèÿ. Îáðàçîâàíèå ìèêðîÿäåð ìîæåò áûòü ñâÿçàíî ñ ïîâûøåíèåì óðîâíÿ ýíäîìóòàãåíåçà â îðãàíèçìå áîëüíûõ è, âåðîÿòíî, âûïîëíÿåò îïðå-äåëåííóþ ðîëü â ðåãóëÿöèè ïðîöåññà àïîïòîçà [25]. Ìèêðîÿäåðíûé òåñò â ëèìôîöèòàõ ïåðèôåðè-÷åñêîé êðîâè (ËÏÊ) â ïîñëåäíèå äåñÿòèëåòèÿ èñ-ïîëüçîâàëè äëÿ îöåíêè âëèÿíèÿ ãåíîòîêñè÷åñêèõ àãåíòîâ íà ÷åëîâåêà. Êîíöåïòóàëüíîé îñíîâîé ýòî-ãî ïîäõîäà ñòàëà ãèïîòåçà, ÷òî ãåíåòè÷åñêèå ïî-âðåæäåíèÿ â ËÏÊ îòðàæàþò êðèòè÷åñêèå ñîáûòèÿ
Figures to article by O. A. Kovalova et al.
à á â
Ðèñ. 8. Àìèòîç ñ îáðàçîâàíè-åì ìåæúÿäåðíîãî ìîñòà:à, â– ÿäðà ñ ðàçíîé ñòåïåíüþ ðàñõîæäåíèÿ;á– ÿäðî ñ íà-÷àëüíîé ñòàäèåé îáðàçîâà-íèÿ âòîðîãî ìîñòà. Êëåòî÷-íàÿ ëèíèÿ À-549.´1000
à á â
Ðèñ. 9. Àìèòîç ñ ôîðìèðîâà-íèåì ìèêðîÿäåð:à,á– êëå-òî÷íàÿ ëèíèÿ À-549;â– êëå-òî÷íàÿ ëèíèÿ ÌÊÑ.´1000
à á â
Ðèñ. 10. Ìèêðîÿäðà â îäíî-ÿäåðíûõ êëåòêàõ:à,á– êëå-òî÷íàÿ ëèíèÿ À-549;â– êëå-òî÷íàÿ ëèíèÿ ÌÊÑ.´1000
à á â
ã ä å
äëÿ êàíöåðîãåííûõ ïðîöåññîâ â òêàíÿõ-ìèøåíÿõ [30]. Ïî äàííûì Êîëþáàåâîé [31], ìèêðîÿäåðíûé òåñò öåëåñîîáðàçíî èñïîëüçîâàòü ïðè îïðåäåëåíèè óðîâíÿ îáëó÷åíèÿ áîëüøèõ ãðóïï ëþäåé â äîçå äî 1 Ãð. Ïðèìåíåíèå â êà÷åñòâå ïîêàçàòåëÿ êëåòîê ñ òðåìÿ è áîëåå ìèêðîÿäðàìè ìîæåò ñòàòü äîïîëíè-òåëüíûì ñâèäåòåëüñòâîì ôàêòà îáëó÷åíèÿ, ïîñêîëü-êó òàêèå ôîðìû àíîìàëèé íå âñòðå÷àþòñÿ ó çäîðî-âûõ ëèö.
Êðîìå òîãî, ó÷åò ìèêðîÿäåð â ËÏÊ ëþäåé øè-ðîêî ïðèìåíÿþò â êà÷åñòâå áèîìàðêåðà îíêîëîãè-÷åñêîãî ðèñêà. Âûñîêàÿ ÷àñòîòà êëåòîê ñ ìèêðîÿä-ðàìè àññîöèèðîâàíà ñ íàëè÷èåì ðàçíûõ îíêîëîãè-÷åñêèõ çàáîëåâàíèé, â òîì ÷èñëå ìî÷åïîëîâîé è æåëóäî÷íî-êèøå÷íîé ñèñòåì [27, 32].
Ìèêðîÿäåðíûé òåñò â äâóÿäåðíûõ êëåòêàõ.
 ìèðîâîé ïðàêòèêå îïðåäåëåíèÿ ãåíåòè÷åñêèõ ïî-âðåæäåíèé ÷àñòî èñïîëüçóþò àíàëèç äâóÿäåðíûõ öèòîêèíåç-áëîêèðîâàííûõ ëèìôîöèòîâ ñ âîçìîæ-íîñòüþ âûÿâëåíèÿ ÷àñòîòû ìèêðîÿäåð, óðîâíÿ êëå-òî÷íîé ãèáåëè è ìèòîòè÷åñêîé äèñôóíêöèè [1, 33].
Èçíà÷àëüíî ìåòîä ñîâåðøåíñòâîâàëñÿ äëÿ èçìå-ðåíèÿ ÷àñòîòû ìèêðîÿäåð, îäíàêî çàòåì ïàðàëëåëü-íî åãî ñòàëè ïðèìåíÿòü äëÿ ó÷åòà õðîìîñîìíûõ ìî-ñòîâ, ÿäåðíûõ àíîìàëèé, íåêðîòè÷åñêèõ, àïîïòè-÷åñêèõ è äåëÿùèõñÿ êëåòîê. Õðîìîñîìíûå ìîñòû ïðîèñõîäÿò îò äèöåíòðè÷åñêèõ õðîìîñîì, ó êîòî-ðûõ öåíòðîìåðû ðàçìåùàþòñÿ íà ïðîòèâîïîëîæíûõ ïîëþñàõ êëåòêè â àíàôàçå, èõ íàëè÷èå ñâèäåòåëüñò-âóþò î õðîìîñîìíûõ ïåðåñòðîéêàõ èëè òåëîìåðíûõ àññîöèàöèÿõ. Ïîâðåæäåíèå õðîìîñîìíîãî ìîñòà ïðèâîäèò ê ôîðìèðîâàíèþ ìèêðîÿäåð.
Ãèáðèäèçàöèÿ ñ èñïîëüçîâàíèåì ôëóîðåñöåíò-íûõ ÄÍÊ-çîíäîâ íà äâóÿäåðôëóîðåñöåíò-íûõ êëåòêàõ ïîçâîëÿåò îïðåäåëÿòü ÷àñòîòó àíåóãåííûõ ñîáûòèé – íåðàñ-õîæäåíèÿ è îòñòàâàíèÿ îòäåëüíûõ õðîìîñîì [2]. Íåîáõîäèìîñòü äåòåêöèè àíåóãåííîé êîìïîíåíòû âîçäåéñòâèÿ ïðîäèêòîâàíà êîëîññàëüíûì ìóòàãåí-íûì ïîòåíöèàëîì àíåóïëîèäèè êàê ãåíåòè÷åñêîãî íàðóøåíèÿ, èãðàþùåãî êëþ÷åâóþ ðîëü â çëîêà÷åñò-âåííîé òðàíñôîðìàöèè êëåòîê è êëîíàëüíîé ýâî-ëþöèè íåîïëàçèè [34].
Ìèêðîÿäåðíûé àíàëèç â öèòîêèíåç-áëîêèðî-âàííûõ êëåòêàõ – ýòî âñåñòîðîííèé òåñò, îí äàåò âîçìîæíîñòü ó÷èòûâàòü âñå êëåòêè, â òîì ÷èñëå íå-êðîòè÷åñêèå è àïîïòè÷åñêèå, à òàêæå êîëè÷åñòâî
ÿäåð íà êëåòêó äëÿ âûÿâëåíèÿ öèòîòîêñè÷åñêîé è ìèòîòè÷åñêîé àêòèâíîñòè [1].
Êàê ïðàâèëî, íàðÿäó ñ ó÷åòîì ìèêðîÿäåð âåäóò ïîäñ÷åò ðàçëè÷íûõ ìîðôîëîãè÷åñêèõ èçìåíåíèé ñòðóêòóðû ÿäðà – âñåõ âèäîâ ÿäåð, èìåþùèõ ôîð-ìó, îòëè÷àþùóþñÿ îò îêðóãëîé èëè îâàëüíîé, è ñî-äåðæàùèõ ðàçëè÷íîãî âèäà ëîïàñòè è «âûáóõà-íèÿ», òàê íàçûâàåìûõ ÿäåðíûõ ïðîòðóçèé.
ßäåðíûå àíîìàëèè. ßäåðíûå ïðîòðóçèè
(pro-trusions, broken eggs, nucleoplasmic bridges, nucleus buds) – ÄÍÊ-ñîäåðæàùèå îáðàçîâàíèÿ, ðàñïîëî-æåííûå âíå ÿäðà, øàðîâèäíîé, íèòåâèäíîé èëè èíîé ôîðìû â öèòîïëàçìå, ÷åòêî îãðàíè÷åííûå îò ÿäðà è ñîåäèíÿþùèåñÿ ñ íèì ïåðåìû÷êîé. Ê ÿäðàì àòèïè÷íîé ôîðìû îòíîñÿò òàêæå êëåòêè ñî ñäâîåí-íûì ÿäðîì (êëåòêè ñ äâóìÿ íåðàçîøåäøèìèñÿ ÿä-ðàìè). Äðóãèå íàçâàíèÿ – ÿäðî ñ ïåðåòÿæêîé, ÿäðî ñ íàñå÷êîé.
Ñðåäè ðàçíîâèäíîñòåé ìîðôîëîãè÷åñêèõ ÿäåð-íûõ àíîìàëèé ìîæíî âûäåëèòü ÿäðà ñ âûðîñòàìè â ïðîñòðàíñòâî öèòîïëàçìû, òàê íàçûâàåìûå «õâîñ-òàòûå ÿäðà» [35] (ðèñ. 11, ñì. âêëåéêó).
Îñíîâíûì ìåõàíèçìîì âîçíèêíîâåíèÿ àòèïè÷-íûõ ÿäåð è ïðîòðóçèé, âîçìîæíî, ÿâëÿåòñÿ íå êëà-ñòîãåííîå, à àíåóãåííîå äåéñòâèå òîêñèêàíòîâ, ñâÿ-çàííîå ñ ïîâðåæäåíèåì áåëêîâ âåðåòåíà äåëåíèÿ êëåòêè [36]. Ñäâîåííûå ÿäðà ìîãóò îáðàçîâûâàòüñÿ â ðåçóëüòàòå íåçàâåðøåííîãî ìèòîçà [37].
ßäåðíûå àíîìàëèè, ïî-âèäèìîìó, óêàçûâàþò íå òîëüêî íà äåãåíåðàòèâíûå ïðîöåññû â êëåòêàõ, â êî-òîðûõ îíè íàáëþäàþòñÿ, íî è íà ôàêò ïðåäøåñòâó-þùèõ õðîìîñîìíûõ àáåððàöèé, âñëåäñòâèå êîòî-ðûõ îíè ñôîðìèðîâàëèñü. Íàïðèìåð, îñíîâíîé ïóë äèöåíòðè÷åñêèõ õðîìîñîì â îáëó÷åííûõin vitroè êóëüòèâèðîâàííûõ ñ äîáàâëåíèåì öèòîõàëàçèíà Á ëèìôîöèòàõ ÷åëîâåêà òðàíñôîðìèðóåòñÿ â ãàíòåëå-âèäíûå ÿäðà [38]. Ïîÿâëåíèå êëåòîê ñ îïðåäåëåí-íûìè âèäàìè ìîðôîëîãè÷åñêèõ àíîìàëèé ÿäåð ìî-æåò áûòü àññîöèèðîâàíî ñ êîëüöåâûìè õðîìîñîìà-ìè, ÿâëÿþùèìèñÿ íàäåæíûìè öèòîãåíåòè÷åñêèìè ìàðêåðàìè ëó÷åâûõ âîçäåéñòâèé [35, 39].
Ðàçëè÷íûå ÿäåðíûå âûðîñòû ñâÿçûâàþò òàêæå ñ ìåõàíèçìàìè èçáàâëåíèÿ êëåòêîé îò èçáûòî÷íî àìï-ëèôèöèðîâàííîé ÄÍÊ. Èçâåñòíî, ÷òî àìïëèôèêà-öèÿ ãåíîâ – ýòî ñòàäèÿ ïðîãðåñèè îïóõîëåâûõ ïðî-öåññîâ â êëåòêå [39].
à á â
Ðèñ. 12. Ïðåæäåâðåìåííàÿ êîíäåíñàöèÿ õðîìîñîì (ÐÑÑ):à– ïîëèïëîèäíàÿ êëåòêà ñ ÐÑÑ;á– íà÷àëüíàÿ ñòàäèÿ êîíäåíñàöèè;â– äè-ïëîèäíàÿ êëåòêà ñ ÐÑÑ. Êëåòî÷íàÿ ëèíèÿ ÌÊÑ.´1000
à á â
Ðèñ. 13. Àïîïòîç:à–â– ðàçíûå ñòåïåíè ðàñïàäà ÿäðà. Êëåòî÷íàÿ ëèíèÿ À-549.´1000
Ê ïîÿâëåíèþ êëåòîê ñ àíîìàëüíî èçìåíåííûìè ÿäðàìè ïðèâîäèò òàêæå âîçäåéñòâèå èîíèçèðóþ-ùåé ðàäèàöèè íà êëåòî÷íûå ïîïóëÿöèè [38], êîòî-ðûå â êëèíè÷åñêîé öèòîëîãèè îòíîñÿò ê ïðîÿâëå-íèþ ëó÷åâûõ ïàòîìîðôîçîâ [40]. Íàëè÷èå ïîâû-øåííîé ÷àñòîòû êëåòîê ñ ÿäåðíûìè ïðîòðóçèÿìè â ëèìôîöèòàõ ëþäåé, ïðîæèâàþùèõ íà çàãðÿçíåí-íûõ ðàäèîíóêëèäàìè òåððèòîðèÿõ, ñ÷èòàåòñÿ íåãà-òèâíûì öèòîãåíåòè÷åñêèì ïîêàçàòåëåì [41].
Ïðåæäåâðåìåííàÿ êîíäåíñàöèÿ õðîìîñîì. Ó
ìëåêîïèòàþùèõ êîíäåíñàöèÿ õðîìîñîì òåñíî ñâÿ-çàíà ñ ðàçðóøåíèåì ÿäåðíîé îáîëî÷êè â ðàííåé ïðîôàçå ìèòîçà è ñ ôîðìèðîâàíèåì âåðåòåíà äåëå-íèÿ.  íåêîòîðûõ ñëó÷àÿõ òàêàÿ êîíäåíñàöèÿ ïðî-èñõîäèò äî âñòóïëåíèÿ êëåòîê â ìèòîç. Ýòîò ôåíî-ìåí ïîëó÷èë íàçâàíèå «ïðåæäåâðåôåíî-ìåííàÿ êîíäåí-ñàöèÿ õðîìîñîì» (ÐÑÑ – premature chromosome con-densation) (ðèñ. 12, ñì. âêëåéêó).
PCC âïåðâûå îïèñàíà â 1970 ãîäó Äæîíñîí è Ðàî ïðè èñïîëüçîâàíèè ìåòîäà ìåæêëåòî÷íîãî ñëè-ÿíèÿ [42]. Áûëî ïîêàçàíî, ÷òî ÐÑÑ ïðîèñõîäèò â òîì ñëó÷àå, êîãäà îäíà èç ñëèâøèõñÿ êëåòîê âñòó-ïàåò â ìèòîç. Ïåðâîíà÷àëüíî ýòî ñîáûòèå ñâÿçûâà-ëè ñ ïðèìåíåíèåì ÓÔ-èíàêòèâèðîâàííîãî âèðóñà Ñåíäàé [42], âïîñëåäñòâèè áûëè ðàçðàáîòàíû ìåòî-äû ñëèÿíèÿ êëåòîê ñ èñïîëüçîâàíèåì õèìè÷åñêèõ âåùåñòâ, íàïðèìåð, ïîëèýòèëåíãëèêîëÿ [43].
Ïðè PCC õðîìîñîìû ìîãóò èìåòü ðàçëè÷íóþ ìîðôîëîãèþ, ÷òî îïðåäåëÿåò ñòàäèþ èíäóöèðîâà-íèÿ êëåòêè. Ïî Ñòåâåíñó [44], ïðè PCC â G2-ôàçå õðîìîñîìû ìîðôîëîãè÷åñêè ñõîäíû ñ íîðìàëüíû-ìè íîðìàëüíû-ìèòîòè÷åñêèíîðìàëüíû-ìè õðîìîñîìàíîðìàëüíû-ìè ñ ñåñòðèíñêèíîðìàëüíû-ìè õðîìàòèäàìè è áåç ïðîáåëîâ, õîòÿ õðîìàòèäû ó íèõ äëèííåå, ÷åì ó ìåòàôàçíûõ õðîìîñîì, ò. å. êàê ó õðîìîñîì â íà÷àëå ïðîôàçû. Ïðè PCC â G1-ôàçå â ðåçóëüòàòå íåçàïóùåííîé ðåïëèêàöèè ÄÍÊ ïðèñóò-ñòâóþò íåñåñòðèíñêèå õðîìàòèäû.  S-ôàçå ïðåæ-äåâðåìåííî êîíäåíñèðîâàííûå õðîìîñîìû âûãëÿ-äÿò, êàê êîíäåíñèðîâàííûå ÷àñòè õðîìàòèíà, ïîÿâ-ëÿþòñÿ òàêæå õðîìîñîìû ñ ìíîæåñòâåííûìè ïðî-áåëàìè, êîëè÷åñòâî êîòîðûõ çàâèñèò îò ñòåïåíè ðå-ïëèêàöèè ÄÍÊ [45].
Ê ïðåæäåâðåìåííîé êîíäåíñàöèè îãðàíè÷åííî-ãî êîëè÷åñòâà õðîìîñîì ìîæåò ïðèâîäèòü ïðèñóò-ñòâèå ìíîãîÿäåðíûõ êëåòîê ñ àñèíõðîííîé ðåïëè-êàöèåé [44].
Âûñîêàÿ ÷àñòîòà âñòðå÷àåìîñòè ñïîíòàííîé ÐÑÑ, òèïè÷íàÿ äëÿ îïóõîëåâûõ êëåòîê, ñâÿçàíà ñ äå-ôåêòîì «òî÷êè ïðîâåðêè» íà ñòàäèè G2/M êëåòî÷-íîãî öèêëà (äèñôóíêöèè áåëêà P53, íàêîïëåíèÿ öèêëèíà B1, àêòèâàöèè cdc2 – öèêëèí-çàâèñèìîé êèíàçû) è ÿâëÿåòñÿ îäíèì èç èñòî÷íèêîâ ãåíåòè÷åñ-êîé ãåòåðîãåííîñòè, èììîðòàëèçàöèè è àäàïòàöèè êëåòî÷íûõ ïîïóëÿöèé ê ðàçíûì óñëîâèÿì õèìèî- è ðàäèîòåðàïèè îïóõîëåé [46, 47].
Ýòà ïàòîëîãèÿ êëåòî÷íîãî öèêëà ìîæåò âûçû-âàòü óâåëè÷åíèå äîëè àíåó- è ïîëèïëîèäíûõ êëå-òîê, ïîÿâëåíèå êîòîðûõ, î÷åâèäíî, ñïîñîáñòâóåò óñ-êîðåíèþ çëîêà÷åñòâåííîé òðàíñôîðìàöèè êëåòîê [48]. Ó÷åò êëåòîê ñ PCC ìîæíî èñïîëüçîâàòü ïðè öè-òîãåíåòè÷åñêîì àíàëèçå ïîêîÿùèõñÿ è ñòàðåþùèõ êëåòîê. Òàêîé àíàëèç èìååò áîëüøîå çíà÷åíèå ïðè îöåíêå êàðèîòèïè÷åñêîãî ðàçíîîáðàçèÿ, íà åãî îñ-íîâå ìîæíî ïðîãíîçèðîâàòü êëèíè÷åñêîå òå÷åíèå ðàçëè÷íûõ çàáîëåâàíèé, âêëþ÷àÿ ìèêðîöåôàëèþ è óìñòâåííóþ îòñòàëîñòü [49], à ñîîòíîøåíèå ìåæäó êëåòêàìè ñ ÐÑÑ è îáùèì ïóëîì äåëÿùèõñÿ êëåòîê ìîæåò ñëóæèòü äîïîëíèòåëüíîé öèòîãåíåòè÷åñêîé õàðàêòåðèñòèêîé ïðè îöåíêå âåðîÿòíîñòè ãåíåòè-÷åñêîé íåñòàáèëüíîñòè êëåòî÷íûõ ïîïóëÿöèé [48].
Îáíàðóæåíî, ÷òî óâåëè÷åíèå ÷àñòîòû âñòðå÷àå-ìîñòè èíòåðôàçíûõ êëåòîê ñ PCC òåñíî ñâÿçàíî ñ êëåòî÷íîé ãèáåëüþ [50]. Ïðåäïîëàãàþò, ÷òî PCC ïðåäøåñòâóåò îäíîìó èç âàðèàíòîâ êëåòî÷íîé ãè-áåëè – «ìèòîòè÷åñêîé êàòàñòðîôå» (ÌÑ), îñóùå-ñòâëÿåìîé ñ ó÷àñòèåì èíûõ, ÷åì ïðè àïîïòîçå, âíóòðèêëåòî÷íûõ ìåõàíèçìîâ [51].
Ìèòîòè÷åñêàÿ êàòàñòðîôà íàáëþäàåòñÿ
ïî-ñëå äåéñòâèÿ ðÿäà ôàêòîðîâ ñòðåññà, âêëþ÷àÿ òåï-ëîâîé øîê, õèìè÷åñêèå àãåíòû, èîíèçèðóþùåå îá-ëó÷åíèå, è õàðàêòåðèçóåòñÿ èçìåíåííîé êëåòî÷íîé ìîðôîëîãèåé [46]. PCC ÿâëÿåòñÿ ðàííèì ýòàïîì ýòî-ãî ïðîöåññà. Äàëåå òàêèå êëåòêè ëèáî íå äîñòèãàþò öèòîêèíåçà, ëèáî äåëÿòñÿ è ïî÷òè ñðàçó ïîñëå ýòîãî ñëèâàþòñÿ, îáðàçóÿ ôèãóðû, òèïè÷íûå äëÿ ãèáåëè ïî òèïó ÌÑ.  òî æå âðåìÿ ïîêàçàíî, ÷òî ÷àñòü êëå-òîê ïåðåæèâàåò ÌÑ è â èõ ïîòîìñòâå ñ âûñîêîé ÷àñ-òîòîé îáíàðóæèâàþòñÿ àíåó- è ïîëèïëîèäû [46, 47].
Ïîíÿòèå «ìèòîòè÷åñêàÿ êàòàñòðîôà» ââåäåíî äëÿ îáîçíà÷åíèÿ ãèáåëè êëåòîê, â êîòîðûõ ïðîÿâëÿ-þòñÿ ïðèçíàêè ïàòîëîãèè ìèòîçà.  ïîñëåäíèå ãî-äû äèñêóòèðóåòñÿ âîïðîñ î òîì, ÷òî ñëåäóåò
âàòü ìèòîòè÷åñêîé êàòàñòðîôîé. Ñîãëàñíî îäíèì ïðåäñòàâëåíèÿì, ÌÑ – ýòî ðåàëèçàöèÿ àïîïòè÷åñ-êîé ïðîãðàììû ñîáñòâåííî â ïðîöåññå ìèòîçà [52]. Ïðè ýòîì ñåãðåãàöèÿ õðîìîñîì îòñóòñòâóåò è êëåò-êà áëîêèðóåòñÿ â îäíîé èç ôàç ìèòîçà. Êàê ïðàâèëî, áëîêèðîâàíèå îñóùåñòâëÿåòñÿ â òàê íàçûâàåìîì Ê-ìèòîçå (êîëõèöèíî-ïîäîáíîì Ê-ìèòîçå), êîãäà â ìè-òîòè÷åñêîé êëåòêå íàðóøåíû îðãàíèçàöèÿ âåðåòåíà è âûñòðàèâàíèå õðîìîñîì â âèäå ìåòàôàçíîé ïëàñ-òèíêè. Äàëåå ïðîèñõîäÿò àêòèâàöèÿ êàñïàç è ïîñëå-äóþùèå äåñòðóêòèâíûå ñîáûòèÿ ïî òèïó àïîïòè÷åñ-êèõ. Ìèòîõîíäðèàëüíûé ïóòü àêòèâàöèè ïðîãðàì-ìû àïîïòîçà ñ÷èòàþò ïðåîáëàäàþùèì ïðè ãèáåëè êëåòîê ñîáñòâåííî â ìèòîçå.
Âòîðûì ïîäòèïîì ÌÑ ÿâëÿåòñÿ ãèáåëü êëåòîê, ïåðåøåäøèõ ïîñëå àíîìàëüíîãî ìèòîçà â ñëåäóþ-ùóþ G1-ôàçó áåç íîðìàëüíîé ñåãðåãàöèè õðîìî-ñîì è îáðàçîâàíèÿ äî÷åðíèõ êëåòîê [53], ò. å. ïîñò-ìèòîòè÷åñêàÿ ãèáåëü ïîëèïëîèäíûõ êëåòîê. Ïðè îáùåé ýóïëîèäíîñòè ïîëèïëîèäíîé êëåòêè åå îò-äåëüíûå ÿäðà ÿâëÿþòñÿ â îñíîâíîì àíåóïëîèäíû-ìè. Äàííûé ïîäòèï ÌÑ ìîæíî íàçâàòü àïîïòîçîì êëåòêè, ïðîøåäøåé ïîëèïëîèäèçèðóþùèé ìèòîç.
Ïðè÷èíîé ÌÑ ñ÷èòàþò èçìåíåíèå ïðîöåññîâ êîíòðîëÿ â êëåòêàõ, â êîòîðûõ ìîãëè ïðîèçîéòè ïî-âðåæäåíèÿ ÄÍÊ èëè íàðóøåíèÿ ñáîðêè âåðåòåíà [52]. Êëþ÷åâûì ìîìåíòîì â áëîêèðîâàíèè êëåòî÷-íîãî öèêëà è â èíäóêöèè â ýòèõ êëåòêàõ àïîïòîçà ÿâëÿåòñÿ ýêñïðåññèÿ áåëêà ð53, âûïîëíÿþùåãî ôóíêöèþ ôàêòîðà òðàíñêðèïöèè äëÿ ð21 – èíãèáè-òîðà G1-ôàçû êëåòî÷íîãî öèêëà – è ðÿäà ïðîàïîï-òè÷åñêèõ áåëêîâ.
Àïîïòîç. Êëåòêè ñ öèòîãåíåòè÷åñêèìè
íàðóøå-íèÿìè ÿâëÿþòñÿ äåôåêòíûìè è ìîãóò óäàëÿòüñÿ èç ïîïóëÿöèè çà ñ÷åò èíòåíñèôèêàöèè àïîïòîçà. Ïðî-ãðàììà àïîïòè÷åñêîé ãèáåëè ñîñòîèò èç ñëåäóþ-ùèõ ãëàâíûõ ýòàïîâ: 1) èíäóêöèÿ ïðîãðàììû àïîï-òîçà; 2) àêòèâàöèÿ ïðîàïîïòè÷åñêèõ áåëêîâ; 3) êàñ-êàä êàñïàç, ðàñùåïëÿþùèõ áåëêè-ìèøåíè; 4) ðàç-ðóøåíèå âíóòðèêëåòî÷íûõ îðãàíåëë èëè èõ ïåðå-ñòðîéêà; 5) ôðàãìåíòàöèÿ êëåòêè íà àïîïòè÷åñêèå òåëüöà; 6) ïîäãîòîâêà êëåòêè è åå ôðàãìåíòîâ ê ôà-ãîöèòîçó ìàêðîôàãàìè èëè ñîñåäíèìè êëåòêàìè.
 çàïóñêå àïîïòîçà ó÷àñòâóþò ðàçíîîáðàçíûå îðãàíåëëû, îäíàêî â ïåðâóþ î÷åðåäü – ýòî ïëàçìà-òè÷åñêàÿ ìåìáðàíà è ìèòîõîíäðèè.
Èíäóêöèÿ àïîïòîçà è ïðîàïîïòè÷åñêèõ áåëêîâ àêòèâèðóþò êàñïàçû (öèñòåèíîâûå ïðîòåàçû). Âû-äåëÿþò èíèöèàòîðíûå è ýôôåêòîðíûå êàñïàçû, ôóíêöèîíèðóþùèå êàê ïðîòåîëèòè÷åñêèå êàñêàäû. Ïîä âëèÿíèåì ýôôåêòîðíûõ êàñïàç ðàçðóøàåòñÿ áîëüøèíñòâî áåëêîâ, ó÷àñòâóþùèõ â ïîääåðæàíèè ãîìåîñòàçà è ðåïàðàöèè êîìïîíåíòîâ êëåòêè, áåë-êîâ – ðåãóëÿòîðîâ êëåòî÷íîãî öèêëà, ñòðóêòóðíûõ áåëêîâ è ò. ä.  ðåçóëüòàòå äåéñòâèÿ ýôôåêòîðíûõ êàñïàç è àêòèâèðîâàííûõ èìè ôåðìåíòîâ (ýíäîíóê-ëåàç, ãåëüçîëèíà è äð.) ðàçðóøàåòñÿ òàêæå âíóòðèÿ-äåðíàÿ ëàìèíà, íàðóøàåòñÿ öåëîñòíîñòü ÄÍÊ, ïðî-èñõîäèò ñïåöèôè÷åñêàÿ êîìïàêòèçàöèÿ õðîìàòèíà, íàáëþäàåòñÿ ðàñïàä ýëåìåíòîâ öèòîñêåëåòà, ìèòî-õîíäðèé, àïïàðàòà Ãîëüäæè, ýíäîïëàçìàòè÷íîãî ðå-òèêóëóìà è ò. ï. Êðîìå êàñïàçíîãî, âûäåëÿþò íåêàñ-ïàçíèé ìåõàíèçì àïîïòè÷åñêîé ñìåðòè, äëÿ êîòîðîé õàðàêòåðíû âûõîä èç ìèòîõîíäðèé è ìèãðàöèÿ â ÿä-ðî ôëàâîïÿä-ðîòåèíà AIF è ýíäîíóêëåàçû G, âûçûâàþ-ùèõ ðàñïàä ÿäåðíîé ÄÍÊ íà êðóïíûå ôðàãìåíòû. Ïðè ýòîì íàáëþäàåòñÿ êîíäåíñàöèÿ õðîìàòèíà è ýêñïîçèöèÿ ôîñôàòèäèëñåðèíà âî âíåøíåì ìîíî-ñëîå ïëàçìàòè÷åñêîé ìåìáðàíû [54].
Ìîðôîëîãè÷åñêèå ïðåâðàùåíèÿ â ïðîöåññå àïîïòîçà âûðàæàþòñÿ â ðàçíîé ñòåïåíè ðàñïàäà âíóòðèêëåòî÷íûõ êîìïîíåíòîâ. Êîíå÷íûìè ýòàïà-ìè àïîïòîçà ÿâëÿþòñÿ óïëîòíåíèå öèòîïëàçìû, ôðàãìåíòàöèÿ ÿäåð è ñàìèõ êëåòîê ñ îáðàçîâàíèåì àïîïòè÷åñêèõ òåëåö, ñîäåðæàùèõ ôðàãìåíòû ÿäåð, ýëåìåíòû àïïàðàòà Ãîëüäæè, ìèòîõîíäðèè è ò. ä. (ðèñ. 13, ñì. âêëåéêó).
×àñòîòà âñòðå÷àåìîñòè àïîïòè÷åñêèõ êëåòîê â êà÷åñòâå åäèíñòâåííîãî ïîêàçàòåëÿ, î÷åâèäíî, íå ìîæåò ñëóæèòü àäåêâàòíûì êðèòåðèåì ãåíîòîêñè-÷åñêèõ ýôôåêòîâ, ïîñêîëüêó îíà êîíòðîëèðóåòñÿ öåëûì àíñàìáëåì ãåíîâ, à òàêæå òêàíåñïåöèôè÷å-ñêîé àêòèâíîñòüþ ìàêðîôàãàëüíûõ êëåòî÷íûõ ýëå-ìåíòîâ, ýëèìèíèðóþùèõ àïîïòè÷åñêèå êëåòêè.
Òàêèì îáðàçîì, èññëåäîâàíèå öèòîìà ïîçâîëÿåò âñåñòîðîííå îöåíèòü ñîñòîÿíèå êëåòî÷íîé ïîïóëÿ-öèè. Îá àíåóãåííûõ ñîáûòèÿõ, òàêèõ êàê ïîâðåæäå-íèå áåëêîâ âåðåòåíà äåëåíèÿ êëåòêè è íàðóøåïîâðåæäå-íèå öèòîòîìèè, ñâèäåòåëüñòâóåò óâåëè÷åíèå ÷àñòîòû êëåòîê ñ ìèêðîÿäðàìè (åñëè ìèêðîÿäðà îáðàçîâàíû îòñòàþùèìè õðîìîñîìàìè), ñ ðàçëè÷íûìè ÿäåðíû-ìè ïðîòðóçèÿÿäåðíû-ìè, àÿäåðíû-ìèòîòè÷åñêèÿäåðíû-ìè ÿäðàÿäåðíû-ìè, ñ äâóìÿ
èëè áîëåå ÿäðàìè, à òàêæå ìíîãîïîëþñíûå ìèòîçû. Êëàñòîãåííûå ïîâðåæäåíèÿ âûðàæàþòñÿ â ôîðìè-ðîâàíèè ìèêðîÿäåð (ïðè ðàçðûâå õðîìîñîì), õðî-ìàòèäíûõ è/èëè õðîìîñîìíûõ ìîñòîâ. Íåîáðàòè-ìûå ñîáûòèÿ – èçìåíåíèå ïðîöåññîâ êîíòðîëÿ â êëåòêå, â êîòîðîé ïðîèçîøëî ïîâðåæäåíèå ÄÍÊ èëè íàðóøåíèå ñáîðêè âåðåòåíà, – ïðèâîäÿò ê ïðîãðàì-ìèðîâàííîé êëåòî÷íîé ãèáåëè. Íåõðîìîñîìíûé àíàëèç ïîçâîëÿåò âûÿâèòü êîìïëåêñ öèòîãåíåòè-÷åñêèõ õàðàêòåðèñòèê â ñîìàòèöèòîãåíåòè-÷åñêèõ êëåòêàõ ìëå-êîïèòàþùèõ, íàèáîëåå èíôîðìàòèâíûé äëÿ êàæäî-ãî îòäåëüíîêàæäî-ãî ñëó÷àÿ.
O. À. Kovalova, N. A. Bezdeneznykh, Y. I. Kudryavets
Nonchromosomal cytogenetic analysis of mammal somatic cells
R. E. Kavetsky Institute of Experimental Pathology, Oncology and Radiobiology
45, Vasylkivska Str., Kyiv, Ukraine, 03022
Summary
The mutational events that take place in mammalian somatic cells in-fluenced with different endogenous and exogenous factors are presen-ted in this review. The nonchromosomal method of research allows ta-king into account the complex cell characteristics without time-consu-ming analysis of the chromosomes as such. As a result, the information can be obtained about the mitotic (phases of mitosis, the number of nuc-lei per cell, micronucnuc-lei, pathology of mitosis) and vital (mitotic index, apoptosis) cell statuses, as well as about the state of chromosomal inte-grity (the presence of nucleoplasmic bridges, nucleus protrusions, chro-mosome fragmentation, micronuclei). Depending on the material stu-died (erythrocytes and lymphocytes of peripheral blood, buccal cells, permanent cell lines etc.), a complex of cytogenetic characteristics can be selected for each case which is the most informative for determina-tion of the mutadetermina-tional spectra in mammalian somatic cells.
Keywords: mutagenesis, somatic cells, cytogenetic analysis, pathology.
Î. À. Êîâàëüîâà, Í. Î. Áåçäºíºæíèõ, Þ. É. Êóäðÿâåöü
Íåõðîìîñîìíèé öèòîãåíåòè÷íèé àíàë³ç ñîìàòè÷íèõ êë³òèí ññàâö³â
Ðåçþìå
 îãëÿä³ ðîçãëÿíóòî ìóòàö³éí³ ïî䳿, ÿê³ â³äáóâàþòüñÿ â ñîìàòè÷-íèõ êë³òèíàõ ññàâö³â ï³ä âïëèâîì ð³çñîìàòè÷-íèõ åíäî- òà åêçîãåíñîìàòè÷-íèõ ôàê-òîð³â. Íåõðîìîñîìíèé ìåòîä äîñë³äæåííÿ äîçâîëÿº âðàõîâóâàòè ñóêóïí³ñòü õàðàêòåðèñòèê êë³òèíè áåç òðóäîì³ñòêîãî àíàë³çó áåçïîñåðåäíüî ñàìèõ õðîìîñîì. Ó ðåçóëüòàò³ ìîæíà îòðèìàòè ³íôîðìàö³þ ïðî ì³òîòè÷íèé (ôàçè ì³òîçó, ê³ëüê³ñòü ÿäåð íà êë³-òèíó, ì³êðîÿäðà, ïàòîëî㳿 ì³òîçó) ³ â³òàëüíèé (ì³òîòè÷íèé ³í-äåêñ, àïîïòîç) ñòàòóñè êë³òèíè, à òàêîæ ïðî ñòàí ö³ë³ñíîñò³ õðîìîñîì (íàÿâí³ñòü íóêëåîïëàçìàòè÷íèõ ìîñò³â, ÿäåðíèõ ïðî-òðóç³é, ôðàãìåíòàö³¿ õðîìîñîì, ì³êðîÿäåð). Çàëåæíî â³ä äîñë³ä-æóâàíîãî ìàòåð³àëó (åðèòðîöèòè ³ ë³ìôîöèòè ïåðèôåð³éíî¿ êðîâ³, êë³òèíè áóêàëüíîãî åï³òåë³þ, ïîñò³éí³ êë³òèíí³ ë³í³¿ òîùî) ìîæíà îáðàòè êîìïëåêñ öèòîãåíåòè÷íèõ õàðàêòåðèñòèê, ÿê³ º íàé³íôîðìàòèâí³øèìè äëÿ êîæíîãî îêðåìîãî âèïàäêó ïðè âèÿâ-ëåíí³ ìóòàö³éíèõ ñïåêòð³â ó ñîìàòè÷íèõ êë³òèíàõ ññàâö³â.
Êëþ÷îâ³ ñëîâà: ìóòàãåíåç, ñîìàòè÷íà êë³òèíà, öèòîãåíåòè÷-íèé àíàë³ç, ïàòîëîã³ÿ.
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Received 12.11.12
