端粒酶反义hTERT基因治疗对卵巢癌细胞生长抑制作用及其对顺铂疗效的影响

Study of increasing sensitivity of CDDP to ovarian carcinoma with telomerase hTERT antisense oligodeoxynuletidem.

目的 :探讨端粒酶反义hTERT对卵巢癌细胞生长抑制作用及对顺铂疗效的影响。方法 :以TRAP PCR ELISA法检测卵巢癌SKOV3、ES 2细胞中端粒酶活性 ;以MTT法及流式细胞术测定反义hTERT对卵巢癌细胞生长的抑制作用、细胞周期阻滞及细胞凋亡的诱导作用及对CDDP疗效的影响。结果 :SKOV3及ES 2细胞中端粒酶活性分别为 2 .17U、2 .2 7U。反义hTERT能降低卵巢癌细胞端粒酶活性 ,以剂量依赖性方式抑制癌细胞生长 ;反义hTERT10 μmol L作用 2 4、72h后G1期细胞所占比例分别为 33 16%、5 3 4 7% ,细胞凋亡发生率分别为 11 66%、13 93% ;而联合应用顺铂时 ,G1期细胞比例分别为 4 2 17%、38 98% ,细胞凋亡发生率分别为 18 66%、2 7 61%。当反义hTERT取 10、2 0、4 0 μmol L时 ,卵巢癌SK OV3、ES 2细胞对顺铂的敏感性分别增加 10 4～ 2 5 5倍、11 7～ 2 0 2倍。结论 :端粒酶反义hTERT基因治疗能通过抑制端粒酶活性、增强细胞周期阻滞及诱导细胞凋亡等途径抑制卵巢癌细胞生长 ,与CDDP合用能增强对癌细胞的杀伤作用。

Objective:To study the influence of the inhibition and the sensitivity of CDDP to ovarian cancer with telomerase hTERT antisense oligodeoxynuletidem.Methods:The activity of telomerase in ovarian carcinoma cell lines SKOV3 and ES-2 was tested by TRAP-PCR-ELISA.The cell growth inhibition,G 1 arrest and apoptosis caused by telomerase hTERT antisense oligodeoxynuletidem were examined by MTT and FAC method.Results:The telomerase activity in SKOV3,ES-2 was 2.17U,2.27U,respectively.The results demonstrated that antisense hTERT inhibited telomerase activity and further inhibited the growth of ovarian cancer cells in a dose-depended way.After reacted with 10μmol/L of antisense hTERT 24h and 72h,the rates of ovarian cancer cells in G 1 stage were 33.16%,53.47%,and the rates of apoptosis were 11.66%,13.93%,respectively.When combined with 5μg CDDP,the rates were 42.17%,38.98%,and 18.66%,27.61%.Under the condition of antisense hTERT 10,20 and 40μmol/L,the sensitivity of SKOV3 and ES-2 to CDDP increases 10.4～25.5 times,11.7～20.2 times,respectively.Conclusions:Telomerase antisense hTERT increases the sensitivity of ovarian carcinoma to CDDP,probably depending on the way which inhibits telomerase activity and induces G 1 stage arrest and apoptosis.