本文選題：非小細胞肺癌（NSCLC） + 多藥耐藥蛋白��； 參考：《大連醫(yī)科大學》2012年碩士論文

【摘要】：目的：為了研究多藥耐藥基因MRP1在非小細胞肺癌中表達的差異，以及觀察siRNA干擾技術(shù)是否能有效抑制非小細胞肺癌耐藥細胞株中M RP1的表達，探討體外高效和特異逆轉(zhuǎn)肺癌耐藥的策略。 方法：運用實時熒光定量PCR、免疫組化法檢測30例NSCLC組織及癌旁組織標本中耐藥基因MRP1mRNA及其蛋白的表達水平，并且以MRP1為靶標設計合成siRNA，直接轉(zhuǎn)染入A549/DDP，轉(zhuǎn)染前后用實時熒光定量PCR檢測MRP1mRNA表達水平，用Western blot檢測MRP1蛋白的表達，MTT法檢測A549/DDP對順鉑耐藥性的變化。 結(jié)果：1.在30例病理組織中，MRP1蛋白在肺癌組織中的陽性率為70%（21/30），癌旁組織中的陽性率為16.67%（5/30），表達有統(tǒng)計學差異(P0.05)；對于癌組織，MRP1蛋白在已經(jīng)化療的病理組織中陽性率為90.9%(10/11)，在未化療的病理組織中陽性率為57.89%(11/19)，，計算兩者有統(tǒng)計學差異(P0.05)，說明MRP1在已化療的組織中高表達。而且PCR結(jié)果顯示MRP1mRNA在癌組織中的表達也高于癌旁組織，經(jīng)過統(tǒng)計學分析有明顯差異(P0.01)。 2.體外實驗中A549/DDP轉(zhuǎn)染MRP1siRNA后,MRP1mRNA表達明顯降低，和轉(zhuǎn)染前相比較有顯著性差異(P0.05)，抑制率達到了78%。轉(zhuǎn)染后MRP1蛋白相對表達強度從之前的0.820±0.037降低為0.435±0.042有明顯差異(P0.05)；MTT結(jié)果顯示轉(zhuǎn)染后A549/DDP對順鉑的敏感性增加了1.43倍。 結(jié)論：1.多藥耐藥基因MRP1在肺癌組織中的高表達,且MRP1蛋白在化療的非小細胞肺癌組織中表達高于未化療組織，說明在非小細胞肺癌患者中存在原發(fā)性耐藥。 2.體外實驗證實了在耐藥細胞轉(zhuǎn)染MRP1siRNA后MRP1的轉(zhuǎn)錄和翻譯都受到了不同程度的抑制，也使其對順鉑的藥物敏感性增強，從一定程度上說明了逆轉(zhuǎn)耐藥。
[Abstract]:Objective: to study the difference of multidrug resistance gene MRP1 expression in non-small cell lung cancer (NSCLC) and whether siRNA interference can effectively inhibit the expression of M RP1 in NSCLC cell lines. Objective: to study the strategy of reversing drug resistance of lung cancer in vitro. Methods: the expression of drug resistance gene MRP1mRNA and its protein in 30 cases of NSCLC and its adjacent tissues were detected by real-time fluorescence quantitative PCR and immunohistochemistry. SiRNAs were designed and synthesized with MRP1 as target and transfected directly into A549 / DDP. The expression of MRP1mRNA was detected by real-time quantitative PCR before and after transfection, and the expression of MRP1 protein was detected by Western blot. The change of cisplatin resistance of A549/DDP to cisplatin was detected by Western blot assay. The result is 1: 1. The positive rate of MRP1 protein was 70% in lung cancer tissue and 16.67% 5 / 30% in paracancerous tissue, and the positive rate of MRP1 protein was 90.9% / 1111% in cancer tissues which had already been treated with chemotherapy, and the positive rate of MRP1 protein was 16.67% 5 / 30% in adjacent tissues, and the positive rate of MRP1 protein in cancer tissues was 90.9% / 1111% in chemotherapeutic pathological tissues, and the positive rate of MRP1 protein was 16.67% 5 / 30% in adjacent tissues of lung cancer. The positive rate of MRP1 was 57.89% and 11 / 19% in pathological tissues. There was a statistical difference between the two groups (P 0.05), which indicated that MRP1 was highly expressed in the tissues that had been treated with chemotherapy. PCR results showed that the expression of MRP1mRNA in cancer tissues was higher than that in paracancerous tissues, and the difference was statistically significant (P 0.01). 2. The expression of MRP1 mRNA in MRP1siRNA transfected with A549/DDP was significantly lower than that before transfection, and the inhibitory rate was 78%. The relative expression intensity of MRP1 protein decreased from 0.820 鹵0.037 to 0.435 鹵0.042 after transfection. The results showed that the sensitivity of A549/DDP to cisplatin increased by 1.43 times after transfection. Conclusion 1. The high expression of multidrug resistance gene MRP1 in lung cancer tissues and the expression of MRP1 protein in non-small cell lung cancer tissues were higher than those in non-small cell lung cancer tissues, indicating that there was primary drug resistance in patients with non-small cell lung cancer. 2. In vitro experiments confirmed that the transcription and translation of MRP1 were inhibited in different degrees after transfection of drug resistant cells into MRP1siRNA, which also enhanced the drug sensitivity to cisplatin, which indicated that the drug resistance was reversed to some extent.
【學位授予單位】：大連醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2012
【分類號】：R734.2;R3416

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