2019 年第 7 期 第 14 卷

长链非编码RNA ENST00000449602基因对胃癌细胞增殖和侵袭及化疗耐药的影响及机制研究

Long-chain non-coding RNA ENST00000449602 gene regulates proliferation, invasion and chemoresistance of gastric cancer cells

作者：孙跃胜潘江华童晓春窦巩昊陈恩德黄立栋胡逸人

英文作者：

单位：325000浙江省温州市人民医院胃肠外科

英文单位：

关键词：胃肿瘤；长链非编码RNAENST00000449602；细胞增殖；侵袭

英文关键词：

• 摘要：

• 【摘要】目的    探讨长链非编码RNA（lncRNA）ENST00000449602基因对胃癌细胞增殖、侵袭及化疗耐药的影响与机制。方法    应用胃癌NU-638细胞株和胃癌组织原代细胞，通过沉默细胞系中lncRNA ENST00000449602表达，分别采用噻唑蓝法和Transwell法检测对胃癌细胞增殖和侵袭能力的影响。流式细胞仪检测过表达lncRNA对顺铂诱导胃癌细胞凋亡的影响。蛋白质印迹法检测过表达lncRNA胃癌细胞组成型光形态建成蛋白1(COP1)、信号传导及转录激活因子3（STAT3）和八聚体结合蛋白4(Oct4)表达水平变化。建立沉默lncRNA胃癌NU-638细胞荷瘤裸鼠模型，观察4周肿瘤体积生长状况。结果    与对照组比较，沉默lncRNA ENST00000449602能明显抑制胃癌NU-638细胞和胃癌组织原代细胞的增殖，细胞抑制率比较［（67.34±0.56）%比（4.34±0.07）%，（59.11±0.65）%比（5.82±0.09）%］，差异均有统计学意义(均P＜0.001)；且能明显抑制细胞侵袭能力［（768±67）个比（1 234±103）个，（656±41）个比（987±83）个］，差异均有统计学意义(均P＜0.05)。与对照组比较，过表达lncRNA可以抑制顺铂诱导的胃癌细胞凋亡（15.15%比21.08%，17.01%比25.22%），差异均有统计学意义（均P＜0.001）。同时，过表达lncRNA能降低E3泛素连接酶COP1表达而提高STAT3和Oct4表达（均P＜0.05）。植入沉默lncRNA ENST00000449602胃癌NU-638细胞的裸鼠4周后肿瘤体积明显小于对照组［（171.0±2.3）mm3比（276.6±2.8）mm3］，差异有统计学意义（P=0.001）。结论    lncRNA ENST00000449602可促进胃癌细胞的增殖、侵袭及顺铂化疗耐药，其机制可能是通过降低COP1蛋白表达提高STAT3和Oct4的表达水平实现。

• 【Abstract】Objective    To investigate the regulation and mechanism of long-chain non-coding RNA(lncRNA) ENST00000449602 gene on proliferation, invasion and chemoresistance of gastric cancer cells. Methods    Human gastric cancer cell line NU-638 and primary cells seperated from gastric cancer tissue were used in the experiment. MTT and Transwell assay were respectively used to detect the effect of silencing lncRNA ENST00000449602 on cell proliferation and invasion. Apoptosis induced by cisplatin of over-expressed lncRNA cancer cells was tested by flow cytometry. Expressions of constitutive photomorphogenic 1 (COP1), signal transducer and activator of transcription 3(STAT3) and octamer-binding protein Oct4 were examined by western blotting. Tumor-bearing nude mouse model of silencing lncRNA NU-638 cells was established and tumor growth was observed after 4 weeks. Results    Silencing lncRNA ENST00000449602 inhibited the proliferation of gastric cancer NU-638 cells and primary cells; cell inhibition rate in silencing lncRNA cells were significantly higher than those in control group［（67.34±0.56）% vs （4.34±0.07）%, （59.11±0.65）% vs （5.82±0.09）%］(both P＜0.001). Silencing lncRNA ENST00000449602 inhibited the invasion of gastric cancer NU-638 cells and primary cells［（768±67） vs （1 234±103）, （656±41） vs （987±83）］(both P＜0.05). Over expression of lncRNA suppressed cisplatin-induced apoptosis of gastric cancer cells（15.15% vs 21.08%, 17.01% vs 25.22%; both P＜0.001）. Expression of E3 ubiquitin ligase protein COP1 decreased significantly in over-expressed lncRNA cancer cells; expressions of STAT3 and Oct4 were higher in over-expressed lncRNA cancer cells than those in control group(all P＜0.05). Tumor size in nude mice of silencing lncRNA NU-638 cells was significantly smaller than that in control group［（171.0±2.3）mm3 vs（276.6±2.8）mm3］(P=0.001). Conclusions    LncRNA ENST00000449602 gene can promote proliferation and invasion of gastric cancer cells. LncRNA ENST00000449602 may increase the sensitivity of gastric cancer cells to cisplatin. The mechanism may be associated with COP1/STAT3/Oct4 signal pathway.