CAJ | 학술논문

目的：研究丙戊酸钠( VPA)联合全反式维甲酸( AT-RA)诱导子宫颈癌细胞衰老作用,并探讨其分子机制。方法实验分为对照组、VPA组、ATRA组、VPA+ ATRA组,采用3 mmol/L VPA、1μmol/L ATRA单独或联合作用于人子宫颈癌HeLa及 SiHa细胞,对照组仅加溶媒；采用 CellTiter 96? AQueous 法检测细胞增殖；β-半乳糖苷酶化学染色法检测细胞衰老；Q-PCR和Western blot法分别检测衰老相关基因P16、P63、hTERT的mRNA和蛋白表达变化。结果 VPA对HeLa及SiHa细胞均有生长抑制作用,与ATRA联合抑制效果明显优于单独用药( P <0．05)；β-半乳糖苷酶染色显示,VPA+ATRA 组衰老细胞比例显著增加,与对照组及VPA组、ATRA 组比较,差异有统计学意义( P <0．05)；Q-PCR和Western blot法结果显示,VPA单独和联合ATRA可上调P16、P63的表达,降低hTERT的表达,且联合用药优于单独用药及对照组( P<0．05)。结论 VPA联合ATRA可抑制子宫颈癌细胞增殖,上调 P16和 P63的表达,下调hTERT的表达,可能是其诱导细胞衰老的分子机制。
목적：연구병무산납( VPA)연합전반식유갑산( AT-RA)유도자궁경암세포쇠로작용,병탐토기분자궤제。방법실험분위대조조、VPA조、ATRA조、VPA+ ATRA조,채용3 mmol/L VPA、1μmol/L ATRA단독혹연합작용우인자궁경암HeLa급 SiHa세포,대조조부가용매；채용 CellTiter 96? AQueous 법검측세포증식；β-반유당감매화학염색법검측세포쇠로；Q-PCR화Western blot법분별검측쇠로상관기인P16、P63、hTERT적mRNA화단백표체변화。결과 VPA대HeLa급SiHa세포균유생장억제작용,여ATRA연합억제효과명현우우단독용약( P <0．05)；β-반유당감매염색현시,VPA+ATRA 조쇠로세포비례현저증가,여대조조급VPA조、ATRA 조비교,차이유통계학의의( P <0．05)；Q-PCR화Western blot법결과현시,VPA단독화연합ATRA가상조P16、P63적표체,강저hTERT적표체,차연합용약우우단독용약급대조조( P<0．05)。결론 VPA연합ATRA가억제자궁경암세포증식,상조 P16화 P63적표체,하조hTERT적표체,가능시기유도세포쇠로적분자궤제。
Objective To study valproate acid( VPA) combined with all-trans retinoic acid( ATRA) induced cervi-cal cancer cells senescence, and explore its mechanism. Methods The experiment is divided into the control group,the VPA group,ATRA and VPA+ATRA groups. HeLa and SiHa cells were treated with combinated use of 3 mmol/L VPA, 1 μmol/L ATRA or respectively. The control group was treated with vehicle only. Using CellTiter 96? Aqueous cell proliferation assay to detect the inhibitory rate of proliferation. Apply β-galactosidase staining method to observe the senescence. The mRNA expressions of P16,P63 and hTERT were detected using Q-PCR and its protein expressions were determined using Western blot. Results The proliferation of HeLa and SiHa cells were significantly inhibited by VPA. The effect of VPA combinated with ATRA was stronger than individual treatment ( P<0. 05 ) . β-galactosidase staining showed that β-galactosidase staining rate was up-regulated after VPA combined with ATRA in cervical cancer HeLa and SiHa cells,the difference was significant compared with control group and individual group ( P<0. 05 ) . Q-PCR and Western blot showed that VPA enhanced the mRNA and protein expres-sion of P16, P63 and reduced the expression of hTERT. The efficacy was more powerful in combination therapy group (P<0. 05). Conclusion VPA combined with ATRA can inhibit the growth of cervical cancer cell lines. The effect of inducing cervical cancer cells senescence may be related with up-regulation of P16 , P63 and down-regulation of hTERT.