摘要
目的探讨富含亮氨酸重复序列免疫球蛋白样蛋白结构域1(LRIG1)在人垂体瘤组织中表达情况,并观察过表达LRIG1后对顺铂(DDP)诱导的垂体瘤细胞增殖、凋亡的影响。方法采用免疫组织化学染色法检测45例垂体瘤患者肿瘤组织及瘤旁组织中的LRIG1表达,分析LRIG1阳性表达与患者临床病理指标的相关性。选取原代分离培养的人垂体瘤细胞,采用脂质体介导的基因转染技术将LRIG1基因过表达质粒p EGFP-N1-LRIG1转染进细胞中,筛选LRIG1过表达的细胞,同时选取转染空质粒p EGFP-N1的细胞作为对照组。两组细胞均经20μg/m L DDP诱导48 h,流式细胞术检测细胞凋亡情况,平板克隆实验检测细胞的增殖能力,Western blot法检测细胞中凋亡相关蛋白BAX、胱天蛋白酶3(caspase-3)和Bcl2的相对表达量。结果垂体瘤患者肿瘤组织中LRIG1阳性率显著低于瘤旁组织,且LRIG1在侵袭性肿瘤表达显著降低。与对照组相比,过表达LRIG1后,DDP诱导的细胞凋亡率显著增加、细胞增殖显著降低、BAX、caspase-3的水平显著增加,而Bcl2水平显著降低。结论过表达LRIG1增加DDP诱导的人垂体瘤细胞凋亡并抑制其增殖。
Objective To investigate the expression of leucine rich repeats and immunoglobulin like domains 1 （LRIG1） in the tumor tissues of human pituitary tumor, and the effects of LRIG1 over-expression on the proliferation and apoptosis of human pituitary tumor cells induced by cisplatin （DDP）. Methods The expression of LRIG1 in the tumor tissues and adjacent tissues of 45 cases with pituitary tumor was detected by immunohistochemistry, and the correlations between the positive expression of LRIG1 and the clinicopathological data of the patients were analyzed. In addition, the isolated human pituitary tumor cells were selected. The LRIG1 gene over-expression plasmid pEGFP-N1-LRIGI was transfected into the cells by liposome-mediated gene transfection. The cells transfected with LRIG1 were screened, and meanwhile, the cells transfected with empty plasmid pEGFP-N1 were selected as a control group. The cells of the two groups were induced by 20 g/mL DDP, and 48 hours later, the cell apoptosis was detected by flow cytometry; the proliferation capacity was tested by plate cloning experiment; and the relative expressions of apoptosis-related proteins Bax, caspase-3 and Bcl2 in the cells were examined by Western blot analysis. Results LRIG1 in the tumor tissues of pituitary tumor was obviously lower than that of the adjacent tissues, and LRIG1 was significantly reduced in the invasive tumor tissues. Compared with the control group, after LRIG1 was over-expressed, the cell apoptosis rate significantly increased, the cell proliferation significantly decreased, and the levels of Bax and caspase-3 increased as well, while the level of Bcl2 decreased remarkably. Condusion Over-expression of LRIG1 increases the apoptosis and inhibits the proliferation of human pituitary tumor cells induced by DDP.
作者
周林裕
包春燕
胡骁
代永庆
包志军
ZHOU Linyu, BAO Chunyan, HU Xiao, DAI Yongqing, BAO Zhijun(3201 Hospital, Medical College, Xi＇an Jiaotong University, Hanzhong 723000, Chin)
出处
《细胞与分子免疫学杂志》
CAS
CSCD
北大核心
2018年第1期9-15,共7页
Chinese Journal of Cellular and Molecular Immunology
