微小RNA181b对胰腺癌细胞增殖和侵袭的影响  

The effects of microRNA-181b on proliferation and invasion of pancreatic cancer cells

作　　者：石英[1] 冯霞[2] 梁红霞[2] 张成武[2] 张军港[2] Shi Ying;Feng Xia;Liang Hongxia;Zhang Chengwu;Zhang Jungang(Department of Obstetrics and Gynecology, Zhejiang Provincial People’s Hospital, Hangzhou 310014, China;Department of Hepatobiliary and Pancreatic Surgery, Zhejiang Provincial People’s Hospital, Hangzhou 310014, China)

机构地区：[1]浙江省人民医院妇产科,杭州310014 [2]浙江省人民医院肝胆胰外科、微创外科,杭州310014

出　　处：《中华实验外科杂志》2019年第4期609-611,共3页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金青年基金(81502482);浙江省医药卫生科研项目(2015KYB026,2017KY193,2017KY210).

摘　　要：目的观察特异性微小RNA181b(miR-181b)对胰腺癌PANC-1细胞增殖和侵袭的影响。方法应用实时荧光定量反转录聚合酶链反应(RT-qPCR)检测3种人胰腺癌细胞株PANC-1、BXPC-3、ASPC-1和正常胰腺HTERT-HPNE细胞中miR-181b表达情况。通过RT-qPCR观察50 nmol/L miR-181b inhibitor对胰腺癌PANC-1细胞miR-181b表达的影响,通过噻唑蓝(MTT)法观察miR-181b inhibitor对胰腺癌PANC-1细胞的增殖影响,通过Transwell法测定miR-181b inhibitor对胰腺癌细胞侵袭的影响,生物信息学预测并通过荧光素酶活性检验miR-181b的靶向基因磷酸酶和张力蛋白同源基因(PTEN),应用RT-qPCR检测miR-181b inhibitor对PTEN,基质金属蛋白酶(MMP)-2和MMP-9表达的影响。结果miR-181b在PANC-1(2.84±0.16)、BXPC-3(1.74±0.18)、ASPC-1(2.21±0.20)细胞中的表达明显高于正常HTERT-HPNE细胞(1.00±0.06)。PANC-1细胞转染miR-181b inhibitor后,细胞增殖无明显变化(P>0.05),侵袭率明显受抑制(P<0.01)。生物信息学预测PTEN可能是miR-181b靶向基因,荧光素酶实验显示转染miR-181b mimics后野生型PTEN组荧光素酶活性下降39%,而突变型差异无统计学意义。转染miR-181b inhibitor后PETN上调(1.85±0.06比1.01±0.08,P<0.05),MMP-2(0.59±0.02比1.01±0.04)和MMP-9(0.66±0.02比0.99±0.07)明显上调(P<0.05)。结论miR-181b可能部分通过靶向PTEN影响MMP-2和MMP-9的表达促进胰腺癌细胞的侵袭性。Objective To investigate the effects of microRNA-181b (miR-181b) on cell proliferation and invasion of pancreatic cancer cells. Methods MiR-181b expression levels in 3 pancreatic cancer cell lines PANC-1, BXPC-3, ASPC-1 and normal pancreatic HTERT-HPNE cells were observed by quantitative reverse transcription-polymerase chain reaction (RT-qPCR). PANC-1 cells were transfected with 50 nmol/L miR-181b inhibitor or negative control using Lipofectamine 2000. After transfection, the expression of miR-181b was measured by RT-qPCR. Cell proliferation was tested by methylthiazol tetrazolium (MTT) assay. Cell invasion was detected by transwell. Bioinformatics prediction and luciferase activity tests were to verify phosphate and tension homology deleted on chromsome ten (PTEN) was the targeted gene of miR-181b. The expression levels of PTEN, matrix metalloproteinase-2 (MMP-2) and matrix metalloproteinase-9 (MMP-9) were measured by RT-qPCR. Results Compared with the HTERT-HPNE cells (1.00±0.06), the expression level of miR-181b in PANC-1 (2.84±0.16), BXPC-3 (1.74±0.18), ASPC-1 (2.21±0.20) was significantly increased. After transfection with 50 nmol/L miR-181b inhibitor, there was no obvious change in PANC-1 cell proliferation (P>0.05), and cell invasive ability was inhibited (P<0.01). Bioinformatics predicted PTEN might be the targeted gene of miR-181b. Luciferase experiments showed that luciferase activity of wild-type PTEN group fell by 39% after transfection of miR-181b mimics, but there was no statistical difference in the mutant group. After transfection with miR-181b inhibitor, PTEN was significantly increased (1.85±0.06 vs. 1.01±0.08, P<0.05), and MMP-2 (0.59±0.02 vs. 1.01±0.04) and MMP-9 (0.66±0.02 vs. 0.99±0.07) were significantly decreased (P<0.05). Conclusion These results demonstrate that miR-181b promotes proliferation and invasion in pancreatic cancer cells, which may be partly related to effecting MMP-2 and MMP-9 expression by PTEN. Therefore, it may be a new target for the biologic therapy for pancr

关 键 词：胰腺癌 微小RNA181b 增殖 侵袭 

分 类 号：R735.9[医药卫生—肿瘤]