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长链非编码RNA ANO1-AS1对食管鳞癌细胞增殖及凋亡的影响

Effect of long non-coding RNA ANO1-AS1 on proliferation and apoptosis of esophageal squamous cell carcinoma cells
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摘要 目的探讨长链非编码RNA(LncRNA)钙离子激活氯离子通道蛋白1反义RNA1(ANO1-AS1)对食管鳞癌(ESCC)细胞增殖和凋亡的影响及其可能的机制。方法在ESCC细胞TE-1和EC109中转染沉默ANO1-AS1慢病毒,qRT-PCR检测其和钙离子激活氯离子通道蛋白1(ANO1)表达情况。CCK-8实验和平板克隆形成实验检测增殖能力;通过LinkedOmics数据库获取ANO1正相关表达基因集并进行通路富集及可能通路的验证;Western blot检测增殖细胞核抗原(PCNA)、抑癌基因P53蛋白、促凋亡蛋白(Bax)、抑凋亡蛋白(Bcl-2)、ANO1蛋白和磷脂酰肌醇-3-激酶/蛋白激酶B(PI3K/Akt)通路相关蛋白的表达情况。结果转染沉默慢病毒后,ANO1-AS1的表达减低(P<0.05);与阴性对照组相比,下调ANO1-AS1后ESCC细胞增殖能力降低(P<0.05),克隆形成率下降(P<0.05);Western blot结果显示,实验组PCNA和Bcl-2蛋白表达降低(P<0.05),P53和Bax蛋白表达升高(P<0.05),通路蛋白PI3K和Akt的磷酸化水平降低(P<0.05)。结论沉默ANO1-AS1可能通过影响PI3K/Akt通路活性从而降低ESCC细胞的增殖能力并促使其发生凋亡。 Objective To investigate the effect of long non-coding RNA(LncRNA)anoctamin 1 antisense RNA-1(ANO1-AS1)on the proliferation and apoptosis of esophageal squamous cell carcinoma(ESCC)cells and its possible mechanisms.Methods Silenced ANO1-AS1 lentivirus was transfected in ESCC cells TE-1 and EC109.Subsequently,the expression levels of ANO1-AS1 and calcium-activated chloride channel protein 1(ANO1)in the cells were detected by qRT-PCR.CCK-8 and colony formation assays were used to detect the proliferation of TE-1 and EC109 cells.ANO1 positively related expressed genes were obtained from the LinkedOmics database and then the gene set was enriched foRpathways and possible pathways were validated.The expression levels of proliferating cell nucleaRantigen(PCNA),P53 protein,apoptosis-related protein(Bax and Bcl-2),ANO1 protein and phosphatidylinositol-3-kinase/protein kinase B(PI3K/Akt)pathway-related protein were assessed by Western blot.Results AfteRtransfection of lentivirus with silent expression function,the expression level of ANO1-AS1 was significantly reduced in TE-1 and EC109 cells(P<0.05);AfteRdown-regulation of ANO1-AS1,compared with the negative control group,the proliferation ability of ESCC cells was reduced(P<0.05)and the rate of clone formation decreased(P<0.05);Western blot results showed that,compared with negative controls,the expression of PCNA decreased,the expression of oncogene P53 protein increased(P<0.05),the expression of proteins(Bax)increased,Bcl-2 decreased and the levels of phosphorylation of the pathway proteins PI3K and Akt decreased(P<0.05).Conclusion Knockdown of ANO1-AS1 can decrease proliferation and promote apoptosis in ESCC,which may be achieved by affecting PI3K/Akt pathway activation.
作者 邵生辉 张健 彭雅琼 向辉 赵敏 谢元茂 郑勇 陈卫刚 Shao Shenghui;Zhang Jian;Peng Yaqiong;Xiang Hui;Zhao Min;Xie Yuanmao;Zheng Yong;Chen Weigang(School of Medicine,Shihezi University,Shihezi 832000;Dept of Gastroenterology,The First Affiliated Hospital,School of Medicine,Shihezi University,Shihezi 832008)
出处 《安徽医科大学学报》 CAS 北大核心 2022年第10期1578-1584,共7页 Acta Universitatis Medicinalis Anhui
基金 中国医学科学院中央级公益性科研院所基本科研业务费专项资金项目(编号:2020-PT330-003) 国家重点研发计划项目(编号:2016YFC1303601) 兵团财政科技计划项目(编号:2020CB002) 石河子大学成果转化与技术推广计划项目(编号:CGZH201704)。
关键词 长链非编码RNA ANO1-AS1 食管鳞癌 增殖 凋亡 long non-coding RNA ANO1-AS1 esophageal squamous cell carcinoma proliferation apoptosis
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