Norrin对胃癌细胞凋亡的抑制作用  

Inhibitory effect of Norrin on apoptosis of gastric cancer cells

作　　者：刘磊[1] 王斌[1] 李砚[1] 刘琴[1] 陈东风[1] 

机构地区：[1]第三军医大学大坪医院野战外科研究所消化内科,重庆400042

出　　处：《第三军医大学学报》2016年第10期1154-1159,共6页Journal of Third Military Medical University

摘　　要：目的研究Norrin蛋白对胃癌细胞凋亡的抑制作用。方法采用免疫组化检测Norrin在151例不同病理分级的胃癌组织中的表达,并进行临床病理分析;Western blot检测不同胃癌细胞株中Norrin的表达情况;随后通过慢病毒转染技术构建Norrin低表达的胃癌细胞株BGC823,并以Western blot检测干扰效果;再以流式细胞术检测Norrin正常表达(Lv-sh-ctrl组)与低表达(Lv-sh-Norrin组)BGC823细胞凋亡的变化,最后使用Western blot检测Mcl-1、Bcl-2、Bax、Cleaved Caspase-3等凋亡相关蛋白的表达。结果 Norrin在胃癌组织中的表达高于癌旁组织,且Norrin的表达高低与胃癌的Lauren分型、分化程度、TNM分期、术后生存率差异有统计学意义(P<0.05),而与性别、年龄、部位、直径、组织学类型差异无统计学意义(P>0.05);在胃癌细胞株中,Norrin在高中分化的SGC7901、AGS细胞中低表达,而在低分化的BGC823细胞中高表达;干扰Norrin基因表达可明显增加胃癌细胞的凋亡,Lv-shNorrin组细胞凋亡率(12.26±0.70)%相比于Lv-sh-ctrl组(7.58±0.68)%明显增加,P<0.05);同时Western blot结果显示,相比于Lv-sh-ctrl组,Lv-sh-Norrin组中抗凋亡蛋白Mcl-1、Bcl-2蛋白表达减少;促凋亡分子Bax、Cleaved Caspase-3蛋白表达增加。结论 Norrin通过抑制Bax、Cleaved Caspase-3蛋白表达,促进Mcl-1、Bcl-2蛋白表达,抑制胃癌细胞的凋亡。Objective To investigate the inhibitory effect of Norrin on apoptosis of gastric cancer cells. Methods The expression of Norrin was detected in 151 gastric cancer tissues with different histological grades by immunohistochemistry,followed by clinic pathological feature analysis. The expression of Norrin in different gastric cell lines was evaluated by Western blot analysis. Gastric cancer cell line BGC823 with low expression of Norrin was constructed by RNA interference using shRNA plasmid. Western blot analysis was applied to confirm the effect of Norrin knockdown. Cell apoptosis in the Norrin normal-expression group（ Lvsh-ctrl group） and Norrin low-expression group（ Lv-sh-Norrin group） was detected by flow cytometry.Finally,Western blot analysis was used to detect the expressions of apoptosis promoting proteins（ Bax and cleaved Caspase-3）,and apoptosis inhibiting proteins（ Mcl-1 and Bcl-2）. Results The expression of Norrin in the gastric tumor tissues was significantly higher than that in the para-tumor tissues. The expression of Norrin was associated with Lauren classification,histological grade,TNM stage and survival rate（ P〈 0. 05）,rather than the gender,age,location,tumor size and histological diagnosis（ P〈 0. 05）. Norrin was highly expressed in poorly differentiated BGC823 cells,but lowly expressed in well or moderately differentiated SGC7901 cells and AGS cells. The apoptotic rate of gastric cancer cells was significantly increased by Norrin knockdown. The apoptosis rate [（ 12. 26 ± 0. 70） %]of the Lv-sh-Norrin group was higher than that [（ 7. 58 ±0. 68） %] of the Lv-sh-ctrl group（ P〈 0. 05）. Western blot analysis showed that Mcl-1 protein and Bcl-2protein were down-regulated in the Lv-sh-Norrin group,while Bax protein and cleaved Caspase-3 were upregulated. Conclusion Norrin inhibits the apoptosis of gastric cancer cells through down-regulation of Bax and cleaved caspase-3 and up-regulation of Mcl-1 and Bcl-2.

关 键 词：Norrin 胃癌 临床病理特征 凋亡 

分 类 号：R341[医药卫生—基础医学] R730.23