DCC基因对卵巢上皮性癌细胞生长及其化疗敏感性的影响

Effects of DCC gene on cell growth and chemosensifivity of ovarian epithelial carcinoma cells

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 study the effects of DCC gene transfection on cell-growth and ehemosensitivity of ovarian epithelial carcinoma cell line HO8910. Methods Recombinant eukaryotie expression vector pcDNA3. 1 （ ＋ ）-DCC containing DCC gene was introduced by lipofectamine transfection reagent into ovarian epithelial carcinoma cell line HO8910 which does not express DCC endogenously. The expression of DCC was detected by RT-PCR and immunocytochemistry. The cell proliferation and the viability rate after different concentrations of eisplatin and paelitaxel were given were assessed by methyl thiazolyl tetrazolium （MTT） assay. Results Exogenous DCC gene had been sueeessfully transferred into HO8910 cells and obtained permanent expression. The growth speed of HO8910-DCC cells was significantly slower than other two groups. There was a significant difference between them （ P 〈 0. 01 ） except at the first day after being planted. There was no difference between the growth speed of HO8910 cells and that of HO8910-Neo cells （P 〉0. 05）. The viability rate of HO8910-DCC cells was significantly lower than other two groups after （ 0. 1 - 5.0 ） peak plasma concentration （ PPC ） of eisplatin and paelitaxel were given （ P 〈 0. 01 ）. The viability rate of HO8910-DCC cells was lower than other two groups after 10. 0 PPC concentration of eisplatin was given （ P 〈 0. 05 ） , but there was no difference between them after 10. 0 PPC concentration of paclitaxel was given （ P 〉 0. 05 ）. The viability rate of HO8910 cells was similar to HO8910- Neo cells after different concentrations of eisplatin and paelitaxel were given （ P 〉 0. 05 ）. Conclusion The DCC gene expression not only inhibits cell growth but also enhances the ehemosensitity of ovarian epithelial carcinoma cell line HO8910.