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背景与目的:实验证明野生型p53基因能增强大多数非小细胞肺癌的化疗敏感性。p53基因的同源体p73与其在结构和功能上都具有高度的相似性。本研究拟探讨p73基因对人肺腺癌细胞株H1299化疗敏感性的影响。方法:通过脂质体介导将p73α基因导入人肺腺癌细胞株H1299,G418筛选获得稳定表达P73α的阳性细胞克隆。Westernblot检测转染细胞中P73α蛋白的表达;MTT法检测细胞存活率,观察转染细胞对阿霉素、顺铂的敏感性变化;采用流式细胞仪和TUNEL法检测.p73646化疗药诱导前后转染细胞凋亡的变化;克隆形成实验观察细胞生物学性状的变化。结果:转染p73α基因的H1299细胞可以稳定高表达P73α蛋白。MTT实验提示顺铂和阿霉素的IC50值分别减少了86.2%和99.2%,转染细胞在原本没有明显抑制和杀伤作用的药物浓度(1.25μmol/L的顺铂或0.05μmol/L的阿霉素)作用下生长明显受到抑制。流式细胞术显示p73α基因转染后顺铂诱导的细胞凋亡率从10.1%升高到38.4%(P<0.01),阿霉素诱导的细胞凋亡率从12.1%升高到49.3%(P<0.01)。克隆形成实验显示p73α基因转染能明显降低化疗后H1299细胞的克隆形成数(P<0.01),对顺铂和阿霉素的化疗增效倍数分别为1.8和2.6倍。结论:转染p73基因可以提高H1299细胞对阿霉素、顺铂等化疗药的敏感性。该作用可?
BACKGROUND & AIM: Wild-type p53 gene has been shown to enhance chemosensitivity in most non-small cell lung cancers. The p53 gene homologue p73 has a high degree of similarity in structure and function. This study was to investigate the effect of p73 gene on chemosensitivity of human lung adenocarcinoma cell line H1299. Methods: The p73α gene was transfected into human lung adenocarcinoma cell lines H1299 and G418 through lipofectamine. The positive clones stably expressing P73α were obtained. The expression of P73αprotein in transfected cells was detected by Western blot, the cell viability was detected by MTT assay, and the sensitivity of transfected cells to doxorubicin and cisplatin was also observed. Flow cytometry and TUNEL assay were used to detect the expression of P733646 The changes of apoptotic cells were observed. The changes of cell biology were observed by clonogenic assay. Results: H1299 cells transfected with p73α gene could stably overexpress P73α protein. MTT assay showed that the IC50 values of cisplatin and doxorubicin decreased by 86.2% and 99.2%, respectively. The transfected cells showed no significant inhibitory and cytotoxic effects (1.25μmol / L cisplatin or 0.05μmol / L ADM) growth was significantly inhibited. Flow cytometry showed that cisplatin-induced apoptosis rate of p73α gene transfected cells increased from 10.1% to 38.4% (P <0.01), and that of doxorubicin-induced apoptosis increased from 12.1% to 49.3% (P < P <0.01). Clone formation experiments showed that the transfection of p73α gene could significantly reduce the number of clonogenic H1299 cells after chemotherapy (P <0.01), and that of cisplatin and doxorubicin were 1.8 and 2.6 times respectively. Conclusion: Transfection of p73 gene can increase the sensitivity of H1299 cells to chemotherapy drugs such as doxorubicin and cisplatin. This effect can?