Huang Wei, Cen Son, Kang Xin-Li, Wang Wei-Fu, Wang Yang, Chen Xiang
Department of Urology, Xiangya Hospital Central South University, Hunan, PR China.
Department of Urology, Hainan Provincial People's Hospital, Hainan, PR China.
Acta Histochem. 2016 Mar;118(2):144-51. doi: 10.1016/j.acthis.2015.12.005. Epub 2015 Dec 31.
To investigate the effect of transforming growth factor-β1 (TGF-β1) on the expression of Fascin1 protein and its impact on cell invasion and metastasis in human renal carcinoma.
Renal tissue slices of 52 cases when undergoing radical nephrectomy were collected to be the observation group, and the normal renal tissues of 23 cases when undergoing nephrectomy due to trauma were collected to be the control group. The expressions of TGF-β1 and Fascin1 were measured by immunohistochemical staining. Human renal carcinoma 786-0 cell line was selected as the study subject. The cells were divided into six groups including NT (no transfection), si-NC (transfection with pGenesil-1-con) si-Fascin1 (transfection with pGen-1-FSCN1) groups, and three corresponding groups: NT, si-NC and si-Fascin1 groups treated with TGF-β1. RT-qPCR, Western-Blot, Transwell, and flow cytometry method were used in this study.
The expressions of TGF-β1 and Fascin1 in the observation group were significantly higher than those in the control group. The expression of TGF-β1 was positively correlated with that of Fascin1. After 24 and 48h of treatment with TGF-β1 (10ng/mL), the invasive and metastatic abilities of the 786-0 cells in the NT and si-NC groups were higher than those before the treatment (P<0.05). Comparing the three groups before TGF-β1 treatment, the invasive and metastatic ability of 786-0 cells in the si-Fascin1 were significantly lower than those in the NT group and si-NC group (P<0.05).
TGF-β1 could induce the expressions of 786-0 Fascin1 mRNA and protein and thus improve the invasive and metastatic ability of human 786-0 renal carcinoma cell.
探讨转化生长因子-β1(TGF-β1)对人肾癌中Fascin1蛋白表达的影响及其对细胞侵袭和转移的作用。
收集52例行根治性肾切除术患者的肾组织切片作为观察组,收集23例行创伤性肾切除术患者的正常肾组织作为对照组。采用免疫组织化学染色法检测TGF-β1和Fascin1的表达。选取人肾癌786-0细胞系作为研究对象。将细胞分为六组,包括未转染组(NT)、阴性对照转染组(si-NC,转染pGenesil-1-con)、Fascin1干扰转染组(si-Fascin1,转染pGen-1-FSCN1),以及三个相应的用TGF-β1处理的组:NT组、si-NC组和si-Fascin1组。本研究采用RT-qPCR、蛋白质免疫印迹法、Transwell小室实验和流式细胞术。
观察组中TGF-β1和Fascin1的表达明显高于对照组。TGF-β1的表达与Fascin1的表达呈正相关。用TGF-β1(10ng/mL)处理24小时和48小时后,NT组和si-NC组中786-0细胞的侵袭和转移能力高于处理前(P<0.05)。在TGF-β1处理前比较三组,si-Fascin1组中786-0细胞的侵袭和转移能力明显低于NT组和si-NC组(P<0.05)。
TGF-β1可诱导786-0细胞Fascin1 mRNA和蛋白的表达,从而提高人786-0肾癌细胞的侵袭和转移能力。