Hsa_circ_0001947对胃癌细胞增殖和凋亡的影响

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摘要［摘要］目的: 探讨环状RNA hsa_circ_0001947(circ_0001947)在胃腺癌细胞和组织中的表达及其对胃癌细胞增殖和凋亡的影响。方法: 通过高通量测序筛选出5例临床胃腺癌、癌旁组织标本中差异表达的5种环状RNA，挑选升高表达最明显的circ_0001947继续研究。实时荧光定量PCR检测circ_0001947在75例临床胃腺癌、癌旁组织以及胃癌BGC-823，HGC-27，MGC-803，SGC-7901细胞株、正常人胃黏膜上皮GES-1细胞株中的表达。将细胞株BGC823、SGC7901各分为两组，设sicirc_0001947组(转染siRNA)和siNC组(转染无关序列)。细胞培养0、12、24、48、72 h后，采用CCK8实验测定细胞活力。采用克隆形成实验测定细胞增殖能力；流式细胞术测定细胞凋亡率，蛋白印迹测定凋亡相关蛋白Cleaved-caspase-3，Bcl-2和Bax表达水平。结果：高通量测序筛选出的胃腺癌组织中差异表达的环状RNA为hsa_circ_0000033，hsa_circ_0000038，hsa_circ_0004916，hsa_circ_0058092，hsa_circ_0001947。circ_0001947在胃腺癌组织中的表达明显高于癌旁组织(P<0.01)，在胃癌BGC-823，HGC-27，MGC-803，SGC-7901细胞株中表达量明显高于正常胃黏膜上皮GES-1细胞株(P均<0.05)。siRNA能够明显抑制BGC823、SGC-7901细胞中circ_0001947表达。si-circ_0001947组BGC823，SGC-7901细胞光密度值明显低于si-NC组(P均<0.01)；sicirc_0001947组BGC823，SGC-7901细胞克隆形成率和细胞凋亡率明显低于siNC组(P均<0.01)。与si-NC组相比，si-circ_0001947组BGC823，SGC7901细胞中Cleavedcaspase-3 及Bax表达明显上调，Bcl2表达明显下调(P均<0.05)。结论： circ_0001947在胃癌细胞和组织中呈高表达，干扰circ_0001947表达可抑制胃癌细胞增殖并诱导其凋亡。

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	陈正威1
	章安伟2
	张垚1
	张烜烽1
	袁海涛1
	步雪峰3

关键词 ： Circ_0001947, 胃腺癌, 细胞增殖, 细胞凋亡

Abstract：[Abstract]Objective: To investigate the expression of circular RNA hsa_circ_0001947(circ_0001947) in gastric cancer cells and tissues and its effect on proliferation and apoptosis of gastric cancer cells. Methods: Five kinds of circular RNA, differentially expressed in 5 cases of gastric carcinoma and paracancerous tissues, were screened by high throughput sequencing. The most obvious circ_0001947 was selected for subsequent study. Realtime fluorescence quantitative reverse transcription PCR was used to detect the expression of circ_0001947 in 75 cases of clinical gastric carcinoma, paracancerous tissues, human gastric cancer BGC-823, HGC-27, MGC-803, SGC-7901 cell lines and normal gastric epithelial GES-1 cell line. The cell lines BGC823 and SGC7901 were divided into two groups：si-circ_0001947 group(transfected with si-RNA) and si-NC group (transfected with irrelevant sequence). Cell viability was determined by CCK8 assay after 0, 12, 24, 48, 72 h of cell culture. Cloning test was used to determine cell proliferation; the cell apoptosis was determined by flow cytometry, Western blotting was used to determine the expression levels of apoptosis-related proteins Cleaved-caspase-3, Bcl-2 and bax. Results: The differentially expressed cyclic RNA in gastric adenocarcinoma tissues screened by high throughput sequencing were hsa_circ_0000033, hsa_circ_0000038, hsa_circ_0004916, hsa_circ_0058092 and hsa_circ_0001947. The expression of circ_0001947 in gastric carcinoma was significantly higher than that in paracancerous tissue (P<0.01). The expression of circ_0001947 in human gastric cancer cell lines BGC-823, HGC-27, MGC-803, SGC-7901 was significantly higher than that in normal gastric epithelial GES1 cell(P<0.05). si-RNA could significantly inhibit the expression of circ_0001947 in BGC-823 and SGC-7901 cells. The absorbance of the BGC823 and SGC7901 cells in the si-circ_0001947 group was significantly lower than that of the si-NC group (P<0.01). The clone formation rate of BGC823 and SGC7901 cells in si-circ_0001947 group was remarkably lower than that in si-NC group (P<0.01). The results of flow cytometry showed that the apoptosis rate of BGC823 and SGC-7901 cells in si-circ_0001947 group was remarkably higher than that in si-NC group(P<0.01). Compared with siNC group, the expression of Cleaved-caspase-3and Bax in BGC-823, SGC-7901 cells of si-circ_0001947 group was significantly upregulated, while the expression of Bcl2 was significantly down-regulated(P<0.05). Conclusion: circ_0001947 was highly expressed in human gastric cancer cells and tissues. Knockdown of circ_0001947 expression could inhibit the proliferation and induce apoptosis of gastric cancer cells.

收稿日期: 2019-10-28

基金资助:［基金项目］国家自然科学基金面上项目(81672999)

通讯作者: 步雪峰(通讯作者)，主任医师，硕士生导师，E-mail：xuefengbu05@163.com

作者简介: 陈正威 (1993—)，男，硕士研究生

引用本文:

陈正威1，章安伟2，张垚1，张烜烽1，袁海涛1，步雪峰3. Hsa_circ_0001947对胃癌细胞增殖和凋亡的影响[J]. 江苏大学学报:医学版, 2020, 30(01): 53-57. CHEN Zheng-wei1, ZHANG An-wei2, ZHANG Yao1, ZHANG Xuan-feng1, YUAN Hai-tao1, BU Xue-feng3. Effect of Hsa_circ_0001947 on proliferation and apoptosis of gastric cancer cells#br# . Journal of Jiangsu University(Medicine Edition), 2020, 30(01): 53-57.

链接本文:

http://zzs.ujs.edu.cn/xbyxb/CN/ 或 http://zzs.ujs.edu.cn/xbyxb/CN/Y2020/V30/I01/53

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