郑州大学学报（医学版）
鄭州大學學報（醫學版）
정주대학학보（의학판）
JOURNAL OF ZHENGZHOU UNIVERSITY(MEDICAL SCIENCES)
2014年
5期
622-625
,共4页
马珊珊%刘静%姚宁%崔渊博%邢衢%王梦然%郭天宇%杨波%关方霞
馬珊珊%劉靜%姚寧%崔淵博%邢衢%王夢然%郭天宇%楊波%關方霞
마산산%류정%요저%최연박%형구%왕몽연%곽천우%양파%관방하
p16%高表达%细胞衰老%调控%293细胞
p16%高錶達%細胞衰老%調控%293細胞
p16%고표체%세포쇠로%조공%293세포
p16%overexpression%cell aging%regulation%293 cell
目的：探讨p16基因高表达对293细胞衰老的影响。方法：实验设正常组、空质粒组和高表达组。 RT-PCR扩增人p16基因，构建p16基因高表达载体，利用脂质体将其转染293细胞。采用CCK-8法和流式细胞术检测p16基因高表达对293细胞增殖和细胞周期的影响，β-半乳糖苷酶染色法检测细胞衰老状态，RT-PCR和Western blot法检测p16、p21 mRNA和蛋白的表达。结果：p16基因高表达可抑制293细胞的增殖，并将细胞阻滞在G0/G1期（F＝158．057，P＜0．001）；高表达组细胞p16和p21在mRNA和蛋白水平的表达明显增加（F＝73．467、54．988、9．923、44．060，P均＜0．05），β-半乳糖苷酶阳性细胞率明显增加（F＝137．266，P＜0．001）。结论：p16基因高表达可抑制293细胞的增殖，将细胞阻滞在G0/G1期，提高细胞中p16和p21的表达，促进细胞衰老。
目的：探討p16基因高錶達對293細胞衰老的影響。方法：實驗設正常組、空質粒組和高錶達組。 RT-PCR擴增人p16基因，構建p16基因高錶達載體，利用脂質體將其轉染293細胞。採用CCK-8法和流式細胞術檢測p16基因高錶達對293細胞增殖和細胞週期的影響，β-半乳糖苷酶染色法檢測細胞衰老狀態，RT-PCR和Western blot法檢測p16、p21 mRNA和蛋白的錶達。結果：p16基因高錶達可抑製293細胞的增殖，併將細胞阻滯在G0/G1期（F＝158．057，P＜0．001）；高錶達組細胞p16和p21在mRNA和蛋白水平的錶達明顯增加（F＝73．467、54．988、9．923、44．060，P均＜0．05），β-半乳糖苷酶暘性細胞率明顯增加（F＝137．266，P＜0．001）。結論：p16基因高錶達可抑製293細胞的增殖，將細胞阻滯在G0/G1期，提高細胞中p16和p21的錶達，促進細胞衰老。
목적：탐토p16기인고표체대293세포쇠로적영향。방법：실험설정상조、공질립조화고표체조。 RT-PCR확증인p16기인，구건p16기인고표체재체，이용지질체장기전염293세포。채용CCK-8법화류식세포술검측p16기인고표체대293세포증식화세포주기적영향，β-반유당감매염색법검측세포쇠로상태，RT-PCR화Western blot법검측p16、p21 mRNA화단백적표체。결과：p16기인고표체가억제293세포적증식，병장세포조체재G0/G1기（F＝158．057，P＜0．001）；고표체조세포p16화p21재mRNA화단백수평적표체명현증가（F＝73．467、54．988、9．923、44．060，P균＜0．05），β-반유당감매양성세포솔명현증가（F＝137．266，P＜0．001）。결론：p16기인고표체가억제293세포적증식，장세포조체재G0/G1기，제고세포중p16화p21적표체，촉진세포쇠로。
Aim:To investigate the regulation of overexpression of p 16 gene on aging of 293 cells.Methods:The ex-periment had 3 groups:control group,empty plasmid group,and overexpression group.The p16 gene was amplified by RT-PCR to construct the p16 gene overexpression vector ,and the vector was transfected into 293 cells.CCK-8 assay and flow cytometry were performed to detect the effect of p 16 gene overexpression on cell proliferation and cell cycle .β-galactosidase staining was used to detect the cell senescence state ,and the expressions of p 16 and p21 in mRNA and protein level were analyzed by RT-PCR and Western blot .Results:p16 gene overexpression could inhibit the proliferation of 293 cells and the cell cycle was arrested in G0/G1 phase(F=158.057,P<0.001).The expressions of p16 and p21 in the overexpression group increased in the mRNA and protein level (F=73.467,54.988,9.923,44.060,P<0.05),and the percentage of cells staining positively for β-galactosidase increased (F=137.266,P <0.001).Conclusion:p16 gene overexpression could inhibit the proliferation of 293 cells,arrest the cell in G0/G1 phase,improve the expression of p16 and p21,and pro-mote the aging .
