论著

microRNA-29b对子宫内膜癌细胞增殖、迁移和侵袭的影响

Effect of microRNA-29b on proliferation,migration and invasion of endometrial cancer cells

:1002-1008
 
目的 探讨微RNA-29b(miR-29b)对子宫内膜癌细胞增殖、迁移和侵袭的影响。方法 子宫内膜癌HEC-1-B细胞分为miR-29b模拟物组(MM组)、miR-29b阻遏物组(MR组)和阴性对照物组(MNC组),分别转染miR-29b拟似物、miR-29b阻遏物和miR-29b阴性对照物,每组设置6个复孔。以实时定量逆转录PCR检测miR-29b表达,以水溶性四氮唑(WST-1)检测miR-29b对HEC-1-B子宫内膜癌细胞增殖的影响,以Transwell小室检测HEC-1-B子宫内膜癌细胞迁移和侵袭的影响,以Western blot法检测磷酸酶张力蛋白同源物(PTEN)-蛋白激酶 B(AKT)通路蛋白表达水平。结果 MNC组、MM组、MR组miR-29b相对表达量分别为(2 032.1±873.4)、(19 272.8±2 087.9)、(472.7±105.6),组间比较差异有统计学意义(P<0.05)。MM组0、3、5、7 d时OD值分别为(0.32±0.06)、(0.53±0.08)、(1.13±0.12)和(1.92±0.14),MNC组0、3、5、7 d时OD值分别为(0.34±0.09)、(0.71±0.08)、(1.67±0.21)和(3.49±0.24),MR组0、3、5、7 d时OD值分别为(0.38±0.09)、(0.84±0.18)、(2.43±0.24)和(5.67±0.15),3组0 d时OD值比较差异无统计学意义(P=0.216),三组3 d、5 d、7 d时OD值比较差异存在统计学意义(P<0.001)。MNC组、MM组和MR组迁移细胞数分别为(403.9±23.8)(102.6±15.7)和(685.7±46.8)个,上述3组侵袭细胞数分别为(82.1±12.7)(38.2±10.6)和(124.6±21.6)个,MM组和MNC组上述指标比较差异均有统计学意义(P<0.05),MR组和MM组上述指标比较差异均有统计学意义(P<0.05)。MNC组、MM组、MR组PTEN蛋白相对表达量分别为(0.25±0.08)、(0.69±0.11)、(0.11±0.05),上述3组p-AKT蛋白相对表达量分别为(0.58±0.10)、(0.13±0.06)和(0.79±0.08),上述3组AKT蛋白相对表达量分别为(0.38±0.09)、(0.37±0.11)和(0.37±0.08),MM组与MNC组PTEN、p-AKT水平比较差异有统计学意义(P<0.05),AKT水平比较差异无统计学意义(P>0.05);MR组与MNC组、MM组PTEN、p-AKT水平比较差异有统计学意义(P<0.05),AKT水平比较差异无统计学意义(P>0.05)。结论 过表达miR-29b对子宫内膜癌细胞增殖、迁移和侵袭具有抑制作用,靶向PTEN-AKT可能是其重要作用途径。
Objective To investigate the effects of microRNA-29b on proliferation,migration and invasion of endometrial cancer cells.Methods The endometrial cancer HEC-1-B cells were divided into micro29b mimetic group(MM group),micro29b repressor group(MR group)and negative control group(MNC group),and the micro29b mimetic,micro29b repressor and micro29b negative control were transfected into each group,six compound holes with each group.The real-time quantitative reverse transcription polymerase chain reaction(RT-PCR)was used to detect the expression of mi29b,WST-1 was used to detect the effect of mi29b on the proliferation of HEC-1-B endometrial cancer cells,Transwell chamber was used to detect the migration and invasion of HEC-1-B endometrial cancer cells,and Western blot was used to detect the expression level of PTEN-AKT pathway protein.Results The relative expression levels of microRNA-29b in MNC group,MM group and MR group were(2 032.1±873.4),(19 272.8±2 087.9)and(472.7±105.6),respectively,and there were significant differences between groups(P<0.05).OD values of MM group at 0 d,3 d,5 d and 7 d were(0.32±0.06),(0.53±0.08),(1.13±0.12)and(1.92±0.14)respectively.The OD values of MNC group at 0,3,5 and 7 days were(0.34±0.09),(0.71±0.08),(1.67±0.21)and(3.49±0.24)respectively.The OD values of MR group at 0 d,3 d,5 d and 7 d were(0.38±0.09),(0.84±0.18),(2.43±0.24)and(5.67±0.15)respectively.There was no significant difference in OD value between the three groups on day 0 (P=0.216).There were significant differences in OD value between the three groups on day 3,day 5 and day 7(P<0.001).The number of migrating cells in MNC group,MM group and MR group were(403.9±23.8)cells,(102.6±15.7)cells and(685.7±46.8)cells,respectively.The number of invasive cells in the above three groups were(82.1±12.7)cells,(38.2±10.6)cells and(124.6±21.6)cells.There were significant differences in the above indexes between MM group and MNC group(P<0.05),also between MR group and MM group(P<0.05).The relative expression levels of PTEN protein in MNC group,MM group and MR group were(0.25±0.08),(0.69±0.11)and(0.11±0.05).The relative expression levels of p-AKT protein in the above three groups were(0.58±0.10),(0.13±0.06)and(0.79±0.08).The relative expression levels of AKT protein in the above three groups were(0.38±0.09),(0.37±0.11)and(0.37±0.08),respectively.Compared with MNC group,the levels of PTEN and p-AKT in MM group had statistical significance(P<0.05),but there was no statistical difference in AKT level(P>0.05).Compared with MNC group and MM group,the levels of PTEN and p-AKT in MR group had statistical significance(P<0.05),and there was no statistical difference in AKT level(P>0.05).Conclusions Overexpression of microRNA-29b can inhibit the proliferation,migration and invasion of endometrial cancer cells,and targeting PTEN-AKT may be an important pathway.
论著

叶酸下调子宫内膜癌中NRBP2基因的表达

Folic acid down-regulates NRBP2 gene expression in endometrial cancer

:19-23
 
目的 探究叶酸对子宫内膜癌作用的靶基因。方法 通过转录组测序筛选叶酸作用下子宫内膜癌细胞中的差异基因,生存分析寻找对子宫内膜癌具有生存意义的差异基因,qPCR及western blot检测其在叶酸作用下的表达。结果 转录组测序发现36个差异基因,生存分析发现FMN1,TRIB3,INHBE及NRBP2的表达对子宫内膜癌具有生存意义,qPCR及western blot验证叶酸作用下NRBP2在子宫内膜癌细胞中的表达下调。结论 叶酸下调子宫内膜癌中NRBP2基因的表达,NRBP2可能是叶酸对子宫内膜癌作用的靶标。
Objective To explore the target genes of folic acid on endometrial carcinoma. Methods The differential genes in endometrial cancer cells treated with folic acid were screened by transcriptome sequencing. Survival analysis was used to find the differential genes with survival significance. QPCR and western blot were used to detect their expression under the action of folic acid. Results 36 differential genes were found by transcriptional sequencing. Survival analysis showed that the expression of FMN1,TRIB3,INHBE and NRBP2 had survival significance in endometrial carcinoma. QPCR and western blot confirmed that the expression of NRBP2 in endometrial cancer cells was down-regulated by folic acid. Conclusion Folic acid down-regulates the expression of NRBP2 gene in endometrial carcinoma, and NRBP2 may be the target of the effect of folic acid on endometrial carcinoma.
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