中国癌症杂志 ›› 2020, Vol. 30 ›› Issue (4): 275-283.doi: 10.19401/j.cnki.1007-3639.2020.04.006

• 论著 • 上一篇    下一篇

PRMT5在胃癌中的表达及其对胃癌细胞增殖、凋亡、迁移能力的影响

燕飞虎 1 ,邹 瑞 2 ,王一尧 1 ,王芳芳 1 ,冉浩男 3 ,王 燕 4   

  1. 1. 海南省肿瘤医院中西医结合科,海南 海口 570100 ;
    2. 海南省肿瘤医院肝胆胰外科,海南 海口 570100 ;
    3. 海南省肿瘤医院放疗科,海南 海口 570100 ;
    4. 海南省肿瘤医院血液肿瘤科,海南 海口 570100
  • 出版日期:2020-04-30 发布日期:2020-05-12
  • 通信作者: 燕飞虎 E-mail: a1980we12@163.com

Expression of PRMT5 in gastric cancer and its influences on proliferation, apoptosis and migration ability of gastric cancer cells

YAN Feihu 1 , ZOU Rui 2 , WANG Yiyao 1 , WANG Fangfang 1 , RAN Haonan 3 , WANG Yan 4   

  1. 1. Department of Integrated Traditional Chinese Medicine and Western Medicine, Hainan Tumor Hospital, Haikou 570100, Hainan Province, China; 2. Department of Hepatobiliary and Pancreatic Surgery, Hainan Tumor Hospital, Haikou 570100, Hainan Province, China; 3. Department of Radiotherapy, Hainan Tumor Hospital, Haikou 570100, Hainan Province, China; 4. Department of Hematology and Oncology, Hainan Tumor Hospital, Haikou 570100, Hainan Province, China
  • Online:2020-04-30 Published:2020-05-12
  • Contact: YAN Feihu E-mail: a1980we12@163.com

摘要: 背景与目的:蛋白精氨酸甲基转移酶5(protein arginine methyltransferase 5,PRMT5)是一种能调控细胞周期的酶,对细胞分裂周期有影响,可以通过调控细胞分裂周期影响细胞的增殖与凋亡。探讨PRMT5在胃癌中的表达及对胃癌细胞增殖、凋亡、迁移能力的影响。方法:采用蛋白质印迹法(Western blot)检测2016年5月—2019年1月在海南省肿瘤医院确诊为胃癌并进行手术治疗的患者的胃癌组织及其癌旁组织(距离癌组织>5 cm)标本(共88例)中的PRMT5的表达,分析PRMT5相对表达量与胃癌临床特征的关系;用PRMT5小干扰RNA(PRMT5-siRNA)和阴性对照(siRNA-NC)转染人胃癌细胞系SGC-7901,以未转染的SGC-7901细胞作为对照组。采用实时荧光定量聚合酶链反应(real-time fluorescence quantitative polymerase chain reaction,RTFQ-PCR)和Western blot检测转染后细胞中PRMT5 mRNA表达和蛋白水平;采用细胞计数试剂盒-8(cell counting kit-8,CCK-8)检测细胞增殖;采用流式细胞术(flow cytometry,FCM)检测细胞凋亡;采用划痕实验检测细胞迁移能力;采用Transwell细胞侵袭实验检测细胞侵袭能力;采用Western blot检测cleaved caspase 3、磷脂酰肌醇3激酶(phosphoinositide 3-kinase,PI3K)、蛋白激酶B(protein kinase B,AKT)和磷酸化AKT( phosphorylated-AKT,p-AKT)蛋白表达。结果:PRMT5在胃癌组织中的表达高于癌旁组织(P<0.01);PRMT5表达水平与胃癌的病理学分级、分期、淋巴结转移有关(P均<0.05);转染后siRNA-NC组和对照组PRMT5 mRNA和蛋白表达量、胃癌细胞的增殖率、凋亡率、cleaved caspase 3、PI3K、AKT、p-AKT蛋白表达量、划痕愈合率、细胞侵袭数差异均无统计学意义(P>0.05),转染后siRNA-NC组和对照组PRMT5 mRNA和蛋白表达量、胃癌细胞增殖率、PI3K、p-AKT、AKT蛋白表达量、划痕愈合率、细胞侵袭数均高于PRMT5-siRNA组,而凋亡率、cleaved caspase 3低于PRMT5-siRNA组(P均<0.05)。结论:PRMT5在胃癌组织中表达明显升高,其表达水平与胃癌的发生、发展、转移恶化密切相关,下调PRMT5的表达可明显减弱胃癌细胞的增殖、迁移能力,同时促进胃癌细胞的凋亡。

关键词: 胃癌, 组蛋白精氨酸甲基转移酶5, 细胞增殖, 细胞凋亡, 迁移能力

Abstract: Background and purpose: Protein arginine methyltransferase 5 (PRMT5) is an enzyme that can regulate cell cycle. It can affect cell proliferation and apoptosis by regulating cell division cycle. This study aimed to explore the expression of PRMT5 in gastric cancer and its influences on proliferation, apoptosis and migration ability of gastric cancer cells. Methods: Western blot was performed to detect PRMT5 expression in gastric cancer tissues and adjacent normal tissues from 88 patients with gastric cancer who received surgery in Hainan Tumor Hospital from May 2016 to Jan. 2019. The relationship between relative expression level of PRMT5 and clinical features of gastric cancer was analyzed. PRMT5 small interfering RNA (PRMT5-siRNA) and negative control (siRNA-NC) were transfected into human gastric cancer cell line SGC-7901. The untransfected SGC-7901 cells were enrolled as control group. Real-time fluorescence quantitative polymerase chain reaction (RTFQ-PCR) and Western blot assay were performed to detect the expression levels of PRMT5 mRNA and protein in cells after transfection. Cell proliferation was detected by cell counting kit-8 (CCK-8) method. Apoptosis was detected by flow cytometry. Cell migration ability was detected by scratch assay. Transwell cell invasion assay was performed to detect cell invasion ability. Western blot method was performed to detect expressions of cleaved caspase-3, phosphoinosmde-3-kinase (PI3K), protein kinase B (AKT) and phosphorylated-AKT (p-AKT) protein. Results: The expression of PRMT5 in gastric cancer tissues was higher than that in adjacent tissues (P<0.01). The expression level of PRMT5 was correlated with pathological grading, staging and lymph node metastasis of gastric cancer (P<0.05). After transfection, there was no significant difference in expression levels of PRMT5 mRNA and protein, proliferation rate of gastric cancer cells, apoptotic rate, protein expression levels of cleaved caspase-3, PI3K, p-AKT and AKT, scratch healing rate and number of cell invasion between siRNA-NC group and control group (P>0.05). After transfection, expression levels of PRMT5 mRNA and protein, proliferation rate of gastric cancer cells, protein expression levels of PI3K, p-AKT and AKT, scratch healing rate and number of cell invasion in siRNA-NC group and control group were higher than those in PRMT5-siRNA group, while apoptotic rate and cleaved caspase-3 expression in siRNA-NC group and control group were lower than those in PRMT5-siRNA group (P<0.05). Conclusion: The expression of PRMT5 is significantly increased in gastric cancer tissues. The expression level of PRMT5 is closely related to occurrence and development of gastric cancer and metastasis. Down-regulation of PRMT5 expression can significantly attenuate proliferation and migration ability and promote apoptosis of gastric cancer cells.

Key words: Gastric cancer, Protein arginine methyltransferase 5, Cell proliferation, Apoptosis, Migration ability