Effect of hMLH1 gene on cisplatin sensitivity of drug-resistant human ovarian cancer cell line

舒丹; 曾飚; 何华; 李渊渊; 钟玲

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Effect of hMLH1 gene on cisplatin sensitivity of drug-resistant human ovarian cancer cell line

更新时间：2026-01-09

- Effect of hMLH1 gene on cisplatin sensitivity of drug-resistant human ovarian cancer cell line
- Journal of Army Medical University Vol. 32, Issue 23, Pages: 2527-2529(2010)
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  CLC： R394.2;R73-36;R737.31
- Published：2010
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舒丹,曾飚,何华,李渊渊,钟玲.hMLH1基因对人卵巢癌耐药细胞株顺铂敏感性的研究[J].陆军军医大学学报(原第三军医大学学报),2010,32(23):2527-2529.

　Shu Dan,Zeng Biao,He Hua,Li Yuanyuan,Zhong Ling.Effect of hMLH1 gene on cisplatin sensitivity of drug-resistant human ovarian cancer cell line[J].J Amry Med Univ (J Third Mil Med Univ),2010,32(23):2527-2529.
舒丹,曾飚,何华,李渊渊,钟玲.hMLH1基因对人卵巢癌耐药细胞株顺铂敏感性的研究[J].陆军军医大学学报(原第三军医大学学报),2010,32(23):2527-2529. DOI：

　Shu Dan,Zeng Biao,He Hua,Li Yuanyuan,Zhong Ling.Effect of hMLH1 gene on cisplatin sensitivity of drug-resistant human ovarian cancer cell line[J].J Amry Med Univ (J Third Mil Med Univ),2010,32(23):2527-2529. DOI：

摘要

目的探讨错配修复基因hMLH1对人卵巢癌耐药细胞株SKOV3/DDP顺铂耐药的逆转作用及其机制。方法分别用重组质粒pcDNA3.1-hMLH1和空质粒pcDNA3.1转染SKOV3/DDP细胞。RT-PCR和Western blot检测hMLH1的表达。MTT检测转染前后SKOV3/DDP细胞对顺铂敏感性的变化。经顺铂DDP作用后

流式细胞术和Western blot法检测细胞凋亡情况。结果与对照组相比，瞬时转染24 h后，SKOV3/DDP细胞中hMLH1 mRNA和蛋白的表达水平均显著升高（P0.05）。MTT结果显示，转染细胞的IC

值（13.95±2.45）较未转染的细胞（94.16±5.18）明显减小。流式细胞仪和Western blot检测结果表明，顺铂作用24 h后，转染了重组质粒的耐药细胞的凋亡率提高到（33.33±2.31）％，同时凋亡抑制蛋白Bcl-2的表达受到抑制（1.35±0.39），与对照组相比，差异均有统计学意义（P0.05）。结论 hMLH1基因能增强卵巢癌耐药细胞对顺铂的敏感性

其机制可能与降低该耐药细胞内Bcl-2的表达水平，促进细胞凋亡有关。

Abstract

Objective To study the reversing effect of hMLH1 gene on cisplatin resistance in ovarian cancer cell line SKOV3/DDP and its underlying mechanism. Methods SKOV3/DDP cells were transfected with recombinant eukaryotic expression plasmid pcDNA3.1-hMLH1 and plasmid pcDNA3.1

respectively

by using lipofectamine 2000. Expression of hMLH1 mRNA and protein was detected by RT-PCR and Western blotting

respectively. Chemosensitivity of SKOV3/DDP cells to cisplatin was assayed by MTT assay. KOV3/DDP cell apoptosis was detected by flow cytometr

y (FCM) and Western blotting

respectively

after treatment with cisplatin. Results MTT assay showed the expression level of hMLH1 mRNA and protein was significantly higher （P0.05） and the IC

value（13.95±2.45）for cisplatin was obviously lower in SKOV3/DDP cells 24 h after transient transfection with hMLH1 gene than in controls（94.16±5.18）. FCM and Western blotting showed that the apoptotic rate of SKOV3/DDP cells transfected with the hMLH1 gene was higher (3.33%±2.31％) than that of controls and the expression level of Bcl-2 was lower in SKOV3/DDP cells transfected with the hMLH1 gene （1.35±0.39） than that in controls （P0.05）. Conclusion The hMLH1 gene can increase the chemosensitivity of human ovarian cancer cells to cisplatin

which may be related to the down-regulation of Bcl-2 expression and improvement of cell apoptosis.

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