过表达长链非编码RNA MT1JP抑制视网膜母细胞瘤SO-RB50细胞的增殖、侵袭和体内移植瘤的生长

Overexpression of long non-coding RNA MT1JP in inhibiting proliferation and invasion of retinoblastoma SO-RB50 cells and growth of transplanted tumors

目的探讨过表达长链非编码RNA MT1JP抑制视网膜母细胞瘤SO-RB50细胞的增殖、侵袭和体内移植瘤的生长机制。方法体外培养视网膜母细胞瘤SO-RB50细胞,并分为空白对照组(control组)、转染对照组(Vector组)及过表达组(MT1JP组);采用RT-PCR检测各组MT1JP的表达;克隆形成检测各组SO-RB50细胞生长情况;流式细胞仪检测细胞凋亡情况;Transwell检测细胞侵袭情况;Western blot检测Ki67、VEGF蛋白表达情况。小鼠皮下注射SO-RB50细胞,建立移植瘤模型;RT-PCR检测长链的表达;统计裸鼠生存时间,绘制生存曲线;检测小鼠肿瘤重量;免疫组化检测小鼠肿瘤组织中Ki67和VEGF的表达情况。结果与空白对照组比较,转染对照组的MT1JP表达、单位面积细胞侵袭数目、Ki67、VEGF蛋白表达均无显著变化(P>0.05);与转染对照组比较,过表达组MT1JP的表达、细胞凋亡率显著升高(P>0.05),克隆实验细胞形成率、Ki67、VEGF蛋白表达、单位面积细胞侵袭数目显著降低(P<0.05);小鼠体内实验显示:与空白对照组比较,转染对照组小鼠肿瘤质量、生存期、Ki67、VEGF蛋白表达均无显著差异(P>0.05);相比转染对照组,过表达组小鼠MT1JP表达、小鼠生存率显著提高(P<0.05);肿瘤质量、Ki67、VEGF蛋白表达显著降低(均P<0.05)。结论过表达长链非编码RNA MT1JP可以抑制视网膜母细胞瘤SO-RB50细胞的增殖、侵袭,促进SO-RB50细胞的凋亡并抑制体内移植瘤的生长。

Objective To investigate mechanism of overexpression of long non-coding RNA MT1 JP in inhibiting proliferation and invasion of retinoblastoma SO-RB50 cells and growth of transplanted tumors.Methods The retinoblastoma SO-RB50 cells were cultured in vitro and divided into Blank control group(control),transfection control group(Vector)and overexpression group(MT1 JP).RT-PCR was performed to detect expression of MT1 JP in each group.The growth of SO-RB50 cells was detected by colony formation.The apoptosis was detected by flow cytometry.The cell invasion was detected by Transwell.The expression of Ki67 and VEGF protein was detected by Western blot.SO-RB50 cells were subcutaneously injected into mice.The transplanted tumor model was established.The expression of long chain was detected by RT-PCR.The survival time of nude mice was statistically analyzed.The survival curve was drawn.The tumor weight of mice was detected.The expression of Ki67 and VEGF in mice tumor tissues was detected by immunohistochemistry.Results Compared with the blank control group,there was no significant change in MT1 JP expression,number of cell invasion per unit area,Ki67 or VEGF protein expression in transfection control group(P>0.05).Compared with transfection control group,MT1 JP expression and apoptosis rate in overexpression group were significantly increased(P>0.05),while cell formation rate of cloning experiment,Ki67,VEGF protein expression and number of cell invasion per unit area were significantly decreased(P<0.05).Compared with blank control group,there was no significant difference in tumor mass,survival time,Ki67 or VEGF protein expression in transfection control group(P>0.05).Compared with transfection control group,MT1 JP expression and survival in overexpression group were significantly increased(P<0.05),while tumor mass,Ki67 and VEGF protein expression were significantly decreased(P<0.05).Conclusion Overexpression of long non-coding RNA MT1 JP can inhibit proliferation and invasion of retinoblastoma SO-RB50 cells,promote apoptosis of SO-RB50 cells and inhibit growth of transplanted tumors.