沉默畸胎瘤衍生生长因子-1基因对胰腺癌细胞株PANC1侵袭力的影响

Effects of teratocarcinoma-derived growth factor-1 on invasion of human pancreatic cancer cell line PANC1

目的 探讨TDGF-1基因沉默对胰腺癌细胞株PANC1侵袭力的影响.方法 设计并合成3个（S1、S2、S3）靶向TDGF-1基因的小干扰RNA（small interfering RNA,siRNA）,筛选出沉默效果最好的siRNA.以该siRNA的不同浓度（3.125、6.25、12.5 nmol/L）转染PANC1细胞,并设对照组和脂质体组.采用实时定量PCR和Western blotting检测TDGF-1 mRNA和蛋白表达,以软琼脂集落培养试验和Boyden小室检测癌细胞的锚着不依赖性增殖和侵袭能力,并将转染48 h的细胞接种裸鼠,观察癌细胞的体内侵袭.结果 siRNA转染组细胞的TDGF-1 mRNA和蛋白表达水平呈浓度和时间依赖性下降,且较脂质体组明显降低（P值均＜0.01）.对照组细胞集落形成数和穿膜细胞数分别为19.8±2.2和49.8±2.6;siRNA组呈浓度依赖性明显减少,12.5 nmol/L转染组分别为5.6±1.2和8.1±1.1.对照组、脂质体组和12.5 nmol/L转染组接种4周后裸鼠种植瘤体积分别为（2.228±0.026）cm3、（2.186±0.028）cm3和（0.728±0.023）cm3.对照组和脂质体组种植瘤侵犯周围肌层组织,转染细胞组未见侵犯.结论 采用RNA干扰技术沉默PANC 1细胞的TDGF-1基因表达可抑制癌细胞的侵袭力.

Objective To investigate the effects of the silence of teratocarcinoma-derived growth factor-1 （ TDGF-1 ） gene on invasion of human pancreatic cancer cell. Methods Three small interfering RNA （siRNA） targeting for TDGF-1 genes （S1, S2, S3 ） were designed and established, then the gene with the best silencing effects was screened. Human pancreatic cancer cell line PANC1 were transfected by siRNA with different concentrations （3. 125, 6.25, 12.5 nmoL/L）, the cells without transfection, and simply treated with liposomes were controls. The expressions of mRNA and protein of TDGF-1 were determined by real time PCR and Western blot assay, respectively. The anchorage-independent growth was examined by clon formation in soft agar, and invasion ability was evaluated by boyden chamber model. PANC1 cells with transfection for 48h were injected into the nude mice to evaluate the invasion ability in vivo. Results The expressions of TDGF-1 mRNA and protein of cells transfected by siRNA were decreased in a dose-and time-dependent manner, which were significantly lower than those in liposomes group. Number of colony formation and transmembrane cell were 19.8 ± 2.2 and 49.8 ＋ 2.6 in the control group, and 5.6 ＋ 1.2 and 8. 1 ＋ 1.1 in the 12.5 nmol/L transfection group. The volumes of tumor 4 weeks after transplation in the control group, liposomes group and the 12.5 nmol/L transfection group were （2.228 ± 0.016 ） cm3, （ 2.186 ± 0.028 ）cm3 and （ 0.728 ± 0.023 ）cm3. Conclusions TDGF-1 gene silence could inhibit invasion ability of human pancreatic cancer cell PANC1.