摘要
目的探讨人miRNA-10b在胰腺癌细胞PANC-1中的表达及其对PANC-1生物学功能的影响。方法采用荧光定量PCR检测miRNA-10b在PANC-1中的表达情况。化学合成成熟型人miRNA-10b模拟体瞬时转染至PANC-1后，分别通过MTT法、流式细胞仪、细胞划痕实验和Transwell小室检测对PANC-1增殖、凋亡、迁移和侵袭功能的影响。结果荧光定量PCR显miRNA-10b在PANC-1中的相对表达量为（3．606±0．423）。瞬时转染后，miRNA-10b转染组miRNA-10b表达量明显增多。MTT法和流式细胞仪示miRNA-10b转染组、阴性对照组和空白对照组细胞的增殖活性和早期凋亡率均无明显差异，细胞划痕实验示miRNA-10b转染组划痕间距缩小明显，侵袭实验miRNA-10b转染组PANC-1细胞穿膜数明显多于对照组。结论miRNA-10b在胰腺癌细胞PANC-1中明显高表达。成熟型miRNA-10b转染后能显著提高转染细胞miRNA-10b的表达量，对PANC-1的体外迁移和侵袭能力有显著的促进作用，但对其增殖和早期凋亡无明显影响。
Objective To investigate the expression of human miRNA-10b in human pancreatic carcinoma PANC-1 cell and the effect of miRNA-10b on the biological function of PANC-1 cell. Methods We adopt the real-time quantitative PCR to detect the expression of miRNA-10b in human pancreatic carcinoma PANC1 cell. The synthesized miRNA-10b was transfeeted into PANC-1 cells via lipofectamine-2000. MTT assay was used to study the effect of miRNA-10b on the proliferation of PANC-1 cells. Flow cytometry was performed to detect the effect of miRNA-10b on the rate of early state apoptosis of PANC-1 cells. The metastasis of PANC-1 cells was detected by cell scratch assay. The invasion of PANC-1 cells was investigated by Transwell Chamber assay. Results Real-time PCR showed that the miRNA-10b expression was （ 3.606 ± 0.423 ） folds that of normal pancreas. After transfection, the expression of miRNA-10b in mimics group was more than that of empty group. The transfection of miRNA-10b had no obvious effect on the proliferation and early state apoptosis of PANC-1 cells. Cell scratch assay pointed out that the nick space of mimics group turned narrower from 24h to 48h.Transwell Chamber assay showed the numbers of cells permeating septum in mimics group were more than other groups obviously. Conclusion MiRNA-10b had significantly higher expression in PANC-1 cell. Synthesized mature type human miRNA-10b can effectively enhance miRNA-10b expression after transfection. The successful transfection of miRNA-10b could promote the invasion and metastasis of the PANC-1 cell, but it had no obvious effect on the proliferation and early state apoptosis.
出处
《浙江临床医学》
2016年第7期1200-1202,共3页
Zhejiang Clinical Medical Journal
