本文選題：hMLH1 + hMSH2��； 參考：《中南大學(xué)》2014年碩士論文

【摘要】：目的：探討hMLH1、hMSH2基因甲基化與宮頸癌細胞順鉑耐藥的相關(guān)性。 方法：1.采用甲基化特異性聚合酶鏈反應(yīng)法(MSP)檢測SIHA及SIHA/DDP(SIHA順鉑耐藥株)中hMLH1、hMSH2基因甲基化的表達差異。2. RT-PCR檢測SIHA及SIHA/DDP中hMLH1、hMSH2基因的表達情況。使用不同濃度5-Aza-CdR預(yù)處理SIHA/DDP,應(yīng)用RT-PCR檢測得到其中hMLH1、hMSH2表達最多時5-Aza-CdR所對應(yīng)的濃度。3.應(yīng)用MTT檢測SIHA、SIHA/DDP及加用5-Aza-CdR的SIHA/DDP,在不同順鉑濃度處理48小時后的耐藥指數(shù)。4.流式細胞儀檢測順鉑對SIHA、SIHA/DDP及加用5-Aza-CdR的SIHA/DDP的細胞周期分布情況的影響。 結(jié)果：1. hMLH1基因在人宮頸癌細胞SIHA中為部分甲基化,而在耐藥細胞SIHA/DDP中則全部甲基化；hMSH2基因在SIHA細胞未見甲基化,在耐藥株中可見部分甲基化。2. hMLH1、hMSH2基因在SIHA細胞中的表達高于在耐藥細胞SIHA/DDP中的表達,并且有統(tǒng)計學(xué)意義(P0.05),5-Aza-CdR干預(yù)SIHA/DDP后,hMLH1、 hMSH2基因隨著的5-Aza-CdR濃度的增加表達逐漸增多。當(dāng)5-Aza-CdR為10μmol/L時,兩基因表達最多。3.通過MTT實驗檢測出SIHA/DDP組耐藥指數(shù)為12.82,經(jīng)5-Aza-CdR處理后的SIHA/DDP組耐藥指數(shù)為6.93,其差異有統(tǒng)計學(xué)意義(P0.05)。4.由流式細胞儀檢測可看出經(jīng)5-Aza-CdR處理后的SIHA/DDP在順鉑作用下細胞顯著減少,以G2期細胞最為顯著。 結(jié)論：hMLH1、hMSH2基因的甲基化參與了宮頸癌耐藥的形成及發(fā)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.
【學(xué)位授予單位】：中南大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2014
【分類號】：R737.33

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