hMLH1、hMSH2基因甲基化与宫颈癌细胞顺铂耐药的相关性研究

发布时间：2018-04-25 12:45

本文选题：hMLH1 + hMSH2　；参考：《中南大学》2014年硕士论文

【摘要】：目的：探讨hMLH1、hMSH2基因甲基化与宫颈癌细胞顺铂耐药的相关性。方法：1.采用甲基化特异性聚合酶链反应法(MSP)检测SIHA及SIHA/DDP(SIHA顺铂耐药株)中hMLH1、hMSH2基因甲基化的表达差异。2. RT-PCR检测SIHA及SIHA/DDP中hMLH1、hMSH2基因的表达情况。使用不同浓度5-Aza-CdR预处理SIHA/DDP,应用RT-PCR检测得到其中hMLH1、hMSH2表达最多时5-Aza-CdR所对应的浓度。3.应用MTT检测SIHA、SIHA/DDP及加用5-Aza-CdR的SIHA/DDP,在不同顺铂浓度处理48小时后的耐药指数。4.流式细胞仪检测顺铂对SIHA、SIHA/DDP及加用5-Aza-CdR的SIHA/DDP的细胞周期分布情况的影响。结果：1. hMLH1基因在人宫颈癌细胞SIHA中为部分甲基化,而在耐药细胞SIHA/DDP中则全部甲基化；hMSH2基因在SIHA细胞未见甲基化,在耐药株中可见部分甲基化。2. hMLH1、hMSH2基因在SIHA细胞中的表达高于在耐药细胞SIHA/DDP中的表达,并且有统计学意义(P0.05),5-Aza-CdR干预SIHA/DDP后,hMLH1、 hMSH2基因随着的5-Aza-CdR浓度的增加表达逐渐增多。当5-Aza-CdR为10μmol/L时,两基因表达最多。3.通过MTT实验检测出SIHA/DDP组耐药指数为12.82,经5-Aza-CdR处理后的SIHA/DDP组耐药指数为6.93,其差异有统计学意义(P0.05)。4.由流式细胞仪检测可看出经5-Aza-CdR处理后的SIHA/DDP在顺铂作用下细胞显著减少,以G2期细胞最为显著。结论：hMLH1、hMSH2基因的甲基化参与了宫颈癌耐药的形成及发展。
[Abstract]:Aim: to investigate the correlation between methylation of hMLH1hMSH2 gene and cisplatin resistance in cervical cancer cells. Method 1: 1. Methylation specific polymerase chain reaction was used to detect the methylation of hMLH1hMSH2 gene in SIHA and SIHA/DDP(SIHA cisplatin resistant strains. The expression of hMLH1 hMSH2 gene in SIHA and SIHA/DDP was detected by RT-PCR. SIHA / DDP was pretreated with different concentrations of 5-Aza-CdR, and the corresponding concentration of 5-Aza-CdR was obtained when hMLH1hMSH2 expression was the highest by RT-PCR detection. MTT was used to detect SIHA / DDP and SIHA / DDP added with 5-Aza-CdR. After 48 hours of treatment with different concentrations of cisplatin, the drug resistance index was 4. 4. The effect of cisplatin on the cell cycle distribution of SIHAN SIHA / DDP and SIHA/DDP with 5-Aza-CdR was detected by flow cytometry. Results 1. HMLH1 gene was partially methylated in human cervical cancer cell line SIHA, while all methylated hMSH2 gene was not methylated in SIHA cell line SIHA/DDP. The expression of methyl.2hMLH1hMSH2 gene in SIHA cells was higher than that in SIHA/DDP cells, and the expression of hMLH1hMSH2 gene in SIHA/DDP cells increased with the increase of 5-Aza-CdR concentration. When 5-Aza-CdR was 10 渭 mol/L, the two genes expressed the most. 3. MTT test showed that the drug resistance index of SIHA/DDP group was 12.82, and that of SIHA/DDP treated with 5-Aza-CdR was 6.93. The difference was statistically significant (P 0.05). Flow cytometry analysis showed that SIHA/DDP treated with 5-Aza-CdR decreased significantly under the action of cisplatin, especially in G2 phase. Conclusion the methylation of the hMLH1hMSH2 gene is involved in the formation and development of drug resistance in cervical cancer.
【学位授予单位】：中南大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R737.33

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