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nm23-H1基因是肿瘤转移抑制基因,转染野生型的nm23-H1基因可以逆转肺癌的恶性表型,我们拟建立转染野生型nm23-H1基因的人大细胞肺癌细胞株L9981-nm23-H1,并初步探讨nm23-H1基因在逆转L9981细胞株恶性表型中的作用。应用基因克隆技术构建质粒载体pLXSN-nm23-H1-EGFP,并建立L9981-nm23-H1细胞株。同时应用Westernblot检测L9981-nm23-H1细胞中nm23-H1蛋白表达。细胞培养及改良的Boyden小室法分别检测L9981-nm23-H1细胞株的体外生物学行为,动物实验检测L9981-nm23-H1细胞株在裸鼠体内的成瘤性及肺部转移能力。结果成功构建了逆转录病毒载体pLXSN-nm23-H1-EGFP;并建立了人大细胞肺癌细胞株L9981-nm23-H1;nm23-H1基因在细胞株L9981-nm23-H1中稳定、高效表达;L9981-nm23-H1细胞体外增殖能力,克隆形成力,体外侵袭力显著降(P<0.01);L9981-nm23-H1裸鼠体内的成瘤性及肺部转移能力显著低于L9981和L9981-pLXSN(P<0.01);nm23-H1基因的抑瘤率82.56%。本研究资料提示转染野生型nm23-H1基因可以逆转人大细胞肺癌细胞株L9981的恶性表型。
The nm23-H1 gene is a tumor metastasis suppressor gene. Transfection of the wild-type nm23-H1 gene can reverse the malignant phenotype of lung cancer. We plan to establish a human large cell lung cancer cell line L9981-nm23-H1 transfected with the wild-type nm23- And to investigate the role of nm23-H1 gene in reversing the malignant phenotype of L9981 cell line. The plasmid vector pLXSN-nm23-H1-EGFP was constructed by gene cloning technique and the L9981-nm23-H1 cell line was established. At the same time, the expression of nm23-H1 protein in L9981-nm23-H1 cells was detected by Western blot. The in vitro biological behavior of L9981-nm23-H1 cell line was detected by cell culture and modified Boyden chamber method. The tumorigenicity and lung metastasis of L9981-nm23-H1 cell line in nude mice were detected by animal experiments. Results The retroviral vector pLXSN-nm23-H1-EGFP was successfully constructed and the human large cell lung cancer cell line L9981-nm23-H1 was established. The nm23-H1 gene was stably and efficiently expressed in L9981-nm23- The ability of tumorigenesis and lung metastasis of L9981-nm23-H1 cells in nude mice was significantly lower than that of L9981 and L9981-pLXSN cells (P <0.01) <0.01). The inhibitory rate of nm23-H1 gene was 82.56%. The data suggest that transfection of wild-type nm23-H1 gene can reverse the malignant phenotype of human large cell lung cancer cell line L9981.