microRNA-100对胰腺癌细胞侵袭和转移能力的影响及机制

Effects and Mechanisms of microRNA-100 on Migration and Invasion in Pancreatic Carcinoma Cells

目的:探讨miR-100对胰腺癌细胞在体外侵袭和迁徙能力的影响及其机制。方法:采用实时荧光定量聚合酶连反应(qRT-PCR)检测miR-100在6株人胰腺癌细胞系及1株人正常胰腺上皮细胞系中的表达,选取表达差异较明显的PANC-1和MIA PaCa-2细胞系进行后续实验;针对miR-100设计miR-100 mimic和空载体(NC),并转染上述两株细胞系中,用PCR检测转染效率;Transwell实验及划痕实验检测miR-100对胰腺癌细胞侵袭转移能力的影响,免疫荧光检测miR-100对胰腺癌细胞骨架的影响,Western blot检测转染细胞后FZD-8的表达。结果:PCR实验显示miR-100在胰腺癌细胞系中低表达,在正常胰腺上皮细胞系中高表达,所设计的miR-100 mimic可以有效地升高PANC-1和MIA PaCa-2细胞系中miR-100的表达;Transwell实验及划痕实验显示转染miR-100之后,能有效的抑制胰腺癌细胞的侵袭及迁移能力(P<0.05);免疫荧光显示转染miR-100后,F-肌动蛋白(F-actin)减少;Western blot结果显示,转染miR-100后,FZD-8的表达减少,胰腺癌细胞的侵袭转移能力明显降低。结论:上调miR-100后,胰腺癌细胞可能通过下调FZD-8的表达来抑制其侵袭转移的能力,提示miR-100有望成为胰腺癌治疗的潜在靶点。

Objective： To investigate the effects and mechanisms of miR-100 on migration and inva- sion in pancreatic carcinoma cells. Methods ： qRT-PCR was used to detect the expression of miR-100 in 6 human pancreatic cancer cell lines and one human normal pancreatic epithelial cell line. Selecting the cell with obvious expression difference PANC-1 and MIA PaCa-2 for further experiments, miR-100 mimic and NC were designed and transfected into the above two cell lines. PCR was used to detect the transfection efficiency. Transwell assay and wound healing assay were used to detect the effects of miR- 100 on pancreatic cancer cells invasion and metastasis ability. The influence of miR-100 on eytoskele- ton was detected by immunofluorescence. The expression of FZD-8 was detected by Western blot. Re- suits： PCR assay showed that miR-lO0 in pancreatic carcinoma cells with low expression in and high expression in normal pancreatic epithehal cell lines. PCR showed that the designed miR-lO0 mimic could effectively improve the expression of miR-100 in PANC-I and MIA PaCa-2 cell lines. Transwell assay and wound healing assay showed that transfected miR-100 could effectively inhibit tile inwlsion and metastasis of pancreatic cancer cells （P 〈 0.05 ）. lmmunofluorescence showed a decrease in F-attin after transfected miR-100. Western blot showed that the expression of FZD-8 was decreased anti the invasion and metastasis of pancreatic cancer cells were significantly decreased alter transfected miR- 100. Conclusion： After improved miR-100, pancreatic cancer cells may inhibit invasion and metastasis by down-regulating the expression of FZD-8. It was suggested that miR-100 could be expected to he a potential target for the treatment of pancreatic cancer.