丙戊酸钠联合全反式维甲酸促进子宫颈癌细胞衰老的发生  

Studies on promoting cellular senescence by VPA combined with ATRA in cervical cancer cells

作　　者：尤青叶 冯定庆[2] 凌斌[1,3] 张红丽[1] 李兵[1] 伍娇娇[1] 赵婷婷[1] 

机构地区：[1]安徽医科大学附属省立医院妇产科,合肥230001 [2]安徽医科大学附属省立医院分子医学重点实验室,合肥230001 [3]卫生部中日友好医院妇产科,北京100086

出　　处：《安徽医科大学学报》2015年第2期135-139,共5页Acta Universitatis Medicinalis Anhui

基　　金：国家自然科学基金(编号:81372777;81372779;81072127)

摘　　要：目的研究丙戊酸钠(VPA)联合全反式维甲酸(ATRA)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He La及Si Ha细胞,对照组仅加溶媒;采用Cell Titer96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、h TERT的mRNA和蛋白表达变化。结果 VPA对He La及Si Ha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);QPCR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h TERT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h TERT的表达,可能是其诱导细胞衰老的分子机制。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.

关 键 词：子宫颈癌 丙戊酸 维甲酸 细胞衰老 

分 类 号：R737.33[医药卫生—肿瘤]