摘要
目的探讨SNHG17在三阴性乳腺癌MDA-MB-231细胞系和组织中的表达及对肿瘤细胞增殖及迁移的影响。方法收集2018年1月至2019年12月深圳市人民医院病理科40例三阴性乳腺癌组织标本,通过碱性磷酸酶原位杂交检测癌和癌旁正常组织中SNHG17的表达情况。构建过表达SNHG17质粒,经一代测序验证质粒。采用荧光定量聚合酶链反应(qRT-PCR)法检测转染SNHG17过表达质粒后过表达组和对照组SNHG17 RNA表达水平。使用CCK8检测过表达SNHG17对MDA-MB-231细胞增殖的影响。通过平板克隆形成实验观察过表达SNHG17对MDA-MB-231细胞克隆能力的影响。进行Transwell小室实验观察过表达SNHG17对肿瘤细胞的迁移能力及侵袭能力的影响。结果原位杂交显示SNHG17在乳腺癌组织表达水平高于癌旁正常组织,差异有统计学意义(P<0.05)。构建SNHG17过表达质粒,经过一代测序验证碱基序列完全匹配。通过qRT-PCR发现,SNHG17过表达组与对照组相比,细胞中SNHG17RNA表达量升高,差异有统计学意义(P<0.05)。CCK8检测表明SNHG17过表达组MDA-MB-231细胞增殖活性较对照组增加,组间比较差异有统计学意义(P<0.05);不同检测时间之间MDA-MB-231细胞增殖活性差异有统计学意义(P<0.05);组间与时间之间存在交互作用,组间差异随时间变化而增加,差异具有统计学意义(P<0.05)。平板克隆形成实验结果显示,SNHG17过表达组克隆细胞数较对照组增加,差异有统计学意义(P<0.05)。进一步Transwell细胞迁移和侵袭实验结果显示,SNHG17过表达组MDA-MB-231细胞迁移和侵袭数目均较对照组增加,差异有统计学意义(P<0.05)。结论SNHG17在MDA-MB-231细胞系及乳腺癌组织中高表达,过表达SNHG17后可促进乳腺癌细胞的增殖、迁移和侵袭。
Objective To investigate the expression of long-chain non-coding RNA(LncRNA)SNHG17 in triple negative breast cancer tissues and cell lines and its effect on the proliferation and migration of breast cancer cells.Methods Tissue samples from 40 patients with triple-negative breast cancer were collected from the Department of Pathology of Shenzhen People’s Hospital from January 2018 to December 2019.The expression of SNHG17 in breast cancer tissues and normal tissues was detected by alkaline phosphatase in situ hybridization.The SNHG17 overexpression plasmid was constructed and the first-generation sequencing was performed to verify the success of the construction.The expression changes of SNHG17 RNA after NC and SNHG17 transfection were detected by qRT-PCR.The effect of SNHG17 overexpression on proliferation and clone formation of breast cancer cells was observed by CCK8 assay and plate clone formation assay.The effect of SNHG17 overexpression on the migration and invasion ability of breast cancer cells was detected by Transwell experiment.Results The expression of SNHG17 in breast cancer tissues was significantly higher than that in paracancerous tissues(P<0.05).Successful construction of the overexpressed plasmid of SNHG17.qRT-PCR showed that the expression level of SNHG17RNA in the transfected SNHG17 overexpressed plasmid group increased compared with that of the control group,and the difference was statistically significant(P<0.05).CCK8 assay showed that the proliferation activity of MDA-MB-231 cells transfected with SNHG17 overexpressed plasmid was significantly higher than that of the control group(P<0.05),and the proliferation activity of MDA-MB-231 cells was significantly different between different detection time(P<0.05).There was an interaction between groups and time,and the difference between groups increased with time,the difference was statistically significant(P<0.05).The results of plate clone formation experiment showed that the number of cloned cells in the SNHG17 overexpression group increased compared with the control group,and the difference was statistically significant(P<0.05).Transwell cell migration and invasion assay showed that the number of mDA-MB-231 cell migration and invasion in SNHG17 overexpression group increased compared with that of the control group,and the difference was statistically significant(P<0.05).Conclusions SNHG17 is highly expressed in MDA-MB-231 cell line and breast cancer tissues,and the overexpression of SNHG17 can promote the proliferation,migration and invasion of breast cancer cells.
作者
许静
高琳
马磊
洪马林
黄净怡
赵盼
XU Jing;GAO Lin;MA Lei(Department of Pathology,the Second Medical College of Ji′nan University,Shenzhen People’s Hospital,Shenzhen 518020,China)
出处
《安徽医学》
2021年第11期1205-1210,共6页
Anhui Medical Journal
基金
广东省基础与应用基础研究基金项目(项目编号:2020A1515111072)
深圳市科技创新委员会基础研究面上项目(项目编号:JCYJ20190806154610953)。
