hMS H2重组质粒对人卵巢癌细胞顺铂敏感性的影响

目的：研究hMSH2重组质粒对卵巢癌顺铂敏感性方面的影响。方法：构建重组真核表达质粒pCAN－hMSH2，通过酶切及测序法对重组质粒进行鉴定。采用脂质体转染技术对卵巢癌耐药细胞SKOV3／DDP进行瞬时转染，对照组为未转染细胞和SKOV3敏感细胞；hMSH2在细胞内的表达变化由Western blotting和RT－PCR进行检测；四甲基偶氮唑蓝（MTT）法对细胞的顺铂敏感性进行检测；耐药细胞的凋亡情况通过Hoechst染色法检测。结果：重组质粒pCAN－hMSH2转染进SKOV3／DDP后，经RT－PCR和Western blotting检测证实转染成功，hMSH2的表达得到增强；MTT结果提示转染后的卵巢癌耐药细胞对顺铂的敏感性明显增强；Ho-echst染色发现，转染后耐药细胞的凋亡明显增强。结论：hMSH2基因转染后能提高其在SKOV3／DDP细胞中的表达，增强SKOV3／DDP细胞对顺铂的敏感性，促进在顺铂作用下的凋亡。

Effects of hMSH2 gene on cisplatin sensitivity of drug-resistant ovarian cancer cells

Objective:To investigate the effect of hMSH2 on cisplatin sensitivity of drug-resistant ovarian cancer cells.Methods:The eukaryotic expresion plasmid pCAN-hMSH2 carrying the coding sequence was constructed and evaluated by enzyme analysis and nucleotide sequencing.The SKOV3/DDP cells were transfected with the recombi-nant plasmid pCAN-hMSH2 and pCAN vector by using Lipofectamine 2000.The expressions of hMSH2 gene were detected by RT -PCR and Western blotting.Chemosensitivity of SKOV3/DDP cells to cisplatin was evaluated by methyl thiazolyl tetrazolium(MTT).The apoptosis of cells was detected by Hoechst dyeing after treatment with cispla-tin.Results:The expression level of hMSH2 gene of SKOV3/DDP cells was increased significantly after transient transfection with pCAN-hMSH2.MTT assay showed the chemosensitivity to cisplatin in SKOV3/DDP cells transfect-ed with hMSH2 was enhanced.Hoechst dyeing showed that the apopototic rate of hMSH2 transfected cells was in-creased.Conclusion:hMSH2 transfection can increase the expression level of hMSH2 gene of SKOV3/DDP cells and enhance the chemosensitivity to cisplatin and improve apoptosis.