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目的:探讨反义脱氧寡核苷酸封闭HSP70基因对体外宫颈癌HeLa细胞增殖、凋亡及化疗敏感性的影响。方法:1)将体外培养的宫颈癌HeLa细胞分为正常对照组(Ctrl组)、反义寡核苷酸处理组(AS组)、正义寡核苷酸处理组(S组)、随机寡核苷酸处理组(R组),每组各5例,分别转染体外培养的宫颈癌HeLa细胞,采用Western免疫印迹检测各组细胞HSP70蛋白表达。2)将顺铂处理体外培养的宫颈癌HeLa细胞分为正常对照组(Ctrl组)、单纯顺铂处理组(Cis组)、反义寡核苷酸+顺铂处理组(AS+Cis组)、正义寡核苷酸+顺铂处理组(S+Cis组)、随机寡核苷酸+顺铂处理组(R+Cis组),每组各8例。采用四甲基偶氮唑蓝光吸收法(methyl thiazolyl tetrazolium,MTT)法检测HeLa细胞的生长抑制率;流式细胞术检测HeLa细胞的凋亡率。结果:1)Ctrl组、AS组、S组、R组HSP70灰度比值分别为1.365±0.187,0.379±0.134,1.403±0.163和1.410±0.158,转染AS组HSP70灰度比值明显下降(P<0.01);2)AS+Cis组生长抑制率和凋亡率明显高于其它各组,差异有统计学意义(P<0.05),S+Cis组和R+Cis组细胞生长抑制率和凋亡率与Cis组相比差异无统计学意义(P>0.05)。结论:HSP70反义脱氧寡核苷酸通过下调HeLa细胞中HSP70蛋白表达,抑制细胞增殖并诱导细胞凋亡。HSP70反义脱氧寡核苷酸与顺铂联合具有协同作用,能提高细胞对顺铂化学治疗的敏感性。
Objective: To investigate the effect of antisense oligodeoxynucleotide-blocked HSP70 gene on the proliferation, apoptosis and chemosensitivity of cervical cancer HeLa cells in vitro. Methods: 1) HeLa cells were divided into normal control group (Ctrl group), antisense oligonucleotide treatment group (AS group), sense oligonucleotide-treated group (S group), random oligonucleotide (R group), 5 in each group. The HeLa cells were transfected with HeLa cells in vitro. Western blotting was used to detect the expression of HSP70 in each group. 2) Cisplatin-treated cervical cancer HeLa cells were divided into normal control group (Ctrl group), cisplatin alone group (Cis group), antisense oligonucleotide + cisplatin group (AS + Cis group) (S + Cis group), random oligonucleotide + cisplatin group (R + Cis group), 8 cases in each group. The growth inhibition rate of HeLa cells was detected by methyl thiazolyl tetrazolium (MTT) method. The apoptosis rate of HeLa cells was detected by flow cytometry. The gray scale ratio of HSP70 in control group, AS group, S group and R group were 1.365 ± 0.187,0.379 ± 0.134,1.403 ± 0.163 and 1.410 ± 0.158 respectively, and the gray value of HSP70 in transfected AS group was significantly decreased (P < 0.01). 2) The growth inhibition rate and apoptosis rate in AS + Cis group were significantly higher than those in other groups (P <0.05). The cell growth inhibition rate and apoptosis in S + Cis group and R + Cis group Compared with Cis group, the difference was not statistically significant (P> 0.05). CONCLUSION: HSP70 antisense oligodeoxynucleotides inhibit HSP70 protein expression in HeLa cells and inhibit cell proliferation and induce apoptosis. HSP70 antisense oligodeoxynucleotide and cisplatin combination has synergistic effect, can improve the cell sensitivity to cisplatin chemotherapy.