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向利军,贺翊峰,鲁才杰,陈伟强,郭亮,季明芳.siRNA干扰Notch1诱导肝癌细胞凋亡的机制研究[J].湖南中医药大学学报,2018,38(3):284-288[点击复制] |
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| siRNA干扰Notch1诱导肝癌细胞凋亡的机制研究 |
| 向利军,贺翊峰,鲁才杰,陈伟强,郭亮,季明芳 |
| (广东医科大学研究生学院, 广东 湛江 524003;广东医科大学附属医院, 广东 湛江 524001;中山市人民医院肝胆外科, 广东 中山 528403;广东医科大学附属中山医院, 广东 中山 528415;中山市肿瘤研究所, 广东 中山 528403) |
| 摘要: |
| 目的 研究Notch1在肝癌组织及细胞中的表达并初步探讨Notch1下调后诱导肝癌细胞调亡的机制。方法 2016年3月至2016年9月于广东医科大学附属医院肝胆外科收集32例肝癌病人样本，利用qRT-PCR方法检测肝癌癌组织及癌旁组织中Notch1基因的表达，免疫组化检测组织中Notch1蛋白表达，siRNA沉默肝癌细胞Notch1表达，流式细胞术检测细胞凋亡情况，Western blot方法检测Notch1、Bcl2和Bax蛋白表达，统计分析Notch1表达水平与肝癌病人临床诊断指标甲胎蛋白（AFP）的相关性。结果 肝癌组织标本中Notch1高表达率为71.9%（23/32），明显高于癌旁组织的28.1%（9/32），差异具有显著统计学意义（P<0.01）；Pearson相关性分析显示，Notch1与AFP存在正相关性（R2=0.3376，P=0.0036）；免疫组化验证Notch1蛋白分别在肝癌癌组织样本中高表达和癌旁组织中低表达；siRNA干扰Notch1基因表达后，镜下发现肝癌细胞4401增殖抑制，流式检测示转染组明显凋亡，蛋白免疫印迹显示凋亡相关蛋白Bcl2蛋白下调、Bax表达上调。结论 Notch1与肝癌的发生、发展相关，下调Notch1可诱导肝癌细胞凋亡，同时Notch1还可作为临床治疗肝癌的潜在新靶点。 |
| 关键词: Notch1 甲胎蛋白 凋亡 肝癌SiRNA |
| DOI：10.3969/j.issn.1674-070X.2018.03.013 |
| 投稿时间：2017-11-15 |
| 基金项目:国家自然科学基金项目（81672098）。 |
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| Mechanism of siRNA Targeting Notch1 on Apoptosis in Hepatocellular Carcinoma Cells |
| XIANG Lijun,HE Yifeng,LU Caijie,CHEN Weiqiang,GUO Liang,JI Mingfang |
| (Postgraduate College of Guangdong Medical University, Zhanjiang, Guangdong 524003, China;Affiliated Hospital of Guangdong Medical University, Zhanjiang, Guangdong 524001, China;Department of Hepatobiliary Surgery, Zhongshan People's Hospital, Zhongshan, Guangdong 528403, China;Affiliated Zhongshan Hospital of Guangdong Medical University, Zhognshan, Guangdong 528415, China;Cancer Institute of Zhongshan, Zhongshan, Guangdong 528403, China) |
| Abstract: |
| Objective To investigate the expression of Notch1 in liver cancer specimens and explain the mechanism of induction of apoptosis by downregulating Notch1 expression. Methods The 32 tissue samples were obtained from Hepatobiliary Surgery of the Affiliated Hospital of Guangdong Medical College during March 2016 to September 2016. The expression of Notch1 gene in hepatocellular carcinoma and adjacent tissues was detected by qRT-PCR. The expression of Notch1 protein was detected by immunohistochemistry. The expression of Notch1, Bcl2 and Bax protein in siRNA was detected by Western blot. The cells were transfected Notch1 siRNA. Apoptosis was detected by flow cytometry. The correlation of Notch 1 expression with the clinical diagnostic index of alpha-fetoprotein (AFP) were revealed by statistical analysis. Results The positive rate of Notch1 in tumor tissues was 71.9% (23/32) and negative rate was 28.1% (9/32), the difference was statistically significant (P<0.01). Pearson correlation analysis showed that there was a positive correlation between Notch1 and AFP(R2=0.3376, P=0.0036). Immunohistochemical results confirmed that Notch1 protein was positive in hepatocellular carcinoma tissues. The data indicated that knockout of Notch1 expression by siRNA would inhibit the proliferation and induce the apoptosis in 4401 cells. In addition, the expression of Bcl2 was attenuated and the expression of Bax was enhanced. Conclusion Notch1 is associated with the development and progression of HCC and the induction of Hepatocellular carcinoma cells apoptosis is driven by down-regulation of Notch1. Notch1 can be used as a potential new target for clinical treatment of hepatocellular carcinoma. |
| Key words: Notch1 alpha fetoprotein apoptosis hepatocellular carcinoma siRNA |
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