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目的研究 DCC 基因对卵巢上皮性癌(卵巢癌)细胞生长及其化疗敏感性的影响。方法采用脂质体转染法将含有 DCC 基因的真核表达载体 pcDNA3.1(+)-DCC 质粒导入卵巢癌细胞系 HO8910细胞中(HO8910-DCC 组),以转染空载体 pcDNA3.1(+)质粒(HO8910-Neo 组)和脂质体(空白对照组)为对照。转染24 h 后,经氨基糖甙类抗生素 G418筛选,RT-PCR 技术以及免疫组化法检测3组细胞 DCC mRNA 及蛋白表达情况;四甲基偶氮唑蓝(MTT)比色法测定3组细胞转染后1～7 d 的细胞生长情况及经梯度浓度[0.1、0.2、1.0、5.0、10.0血浆峰浓度(PPC)]化疗药物顺铂和紫杉醇处理后48 h 的细胞存活率,并绘制细胞生长曲线。结果转染 DCC 基因后,DCC 基因可在HO8910细胞中稳定表达。转染后1～7 d,HO8910-DCC 组细胞生长速度较其他两组细胞明显减慢,除转染后第1天外,分别比较,差异均有统计学意义(P<0.01);空白对照组和 HO8910-Neo 组细胞生长速度比较,差异则无统计学意义(P>0.05)。HO8910-DCC 组细胞经0.1～5.0 PPC 的顺铂和紫杉醇处理后,细胞存活率显著低于空白对照组和 HO8910-Neo 组(P<0.01);经10.0 PPC 的顺铂处理后细胞存活率也明显低于空白对照组和 HO8910-Neo 组(P<0.05);但经10.0 PPC 的紫杉醇处理后细胞存活率与空白对照组和 HO8910-Neo 组相比,差异则无统计学意义(P>0.05)。空白对照组和HO8910-Neo 组细胞经各梯度浓度药物处理后的细胞存活率间比较,差异均无统计学意义(P>0.05)。结论 DCC 基因可明显抑制卵巢癌细胞的生长,并增强卵巢癌细胞对化疗药物的敏感性。
Objective To investigate the effect of DCC gene on the growth and chemosensitivity of ovarian epithelial carcinoma (ovarian cancer) cells. Methods The eukaryotic expression vector pcDNA3.1 (+) - DCC containing DCC gene was transfected into HO8910 ovarian cancer cell line (HO8910-DCC) by lipofection method and transfected into empty vector pcDNA3.1 +) Plasmid (HO8910-Neo group) and liposome (blank control group) as control. The expression of DCC mRNA and protein in three groups of cells was detected by G418 aminoglycoside antibiotics, RT-PCR and immunohistochemistry 24 h after transfection; MTT assay 3 The cell growth and the cell viability at 1 to 7 days after transfection of the group of cells and the cell viability at 48 h after the treatment with cisplatin and paclitaxel at the gradient concentrations [0.1, 0.2, 1.0, 5.0 and 10.0 peak plasma concentrations (PPC)] Draw cell growth curve. Results DCC gene was stably expressed in HO8910 cells after transfection of DCC gene. The cell growth rate of HO8910-DCC group was significantly slower than that of the other two groups at 1 ~ 7 days after transfection, except for the first day after transfection, the difference was statistically significant (P <0.01); the blank control group Compared with HO8910-Neo group, the difference of cell growth rate was not statistically significant (P> 0.05). The cell viability of HO8910-DCC cells treated with cisplatin and paclitaxel of 0.1-5.0 PPC was significantly lower than that of the blank control group and HO8910-Neo group (P <0.01) (P <0.05). However, the cell viability after treatment with paclitaxel 10.0 PPC was not significantly different from that of the blank control group and HO8910-Neo group (P> 0.05) ). There was no significant difference in the cell survival rate between the blank control group and HO8910-Neo group after treatment with each gradient concentration of drug (P> 0.05). Conclusion DCC gene can significantly inhibit the growth of ovarian cancer cells and enhance the sensitivity of ovarian cancer cells to chemotherapy drugs.