可溶性转化生长因子βⅡ型受体对大肠癌生物学行为的影响

Effect of soluble type Ⅱ transforming growth factor(TGF-β)receptor on biological behavior of colorectal carcinoma cell line LoVo

目的探讨可溶性转化生长因子βⅡ型受体(sTGF-βRⅡ)对大肠癌 LoVo 细胞生物学行为的影响。方法采用 DNA 重组技术构建 sTGF-βRⅡ的表达载体 pcDNA3.1(+)/sTGF-βRⅡ,转染人大肠癌 LoVo 细胞,以逆转录-聚合酶链反应(RT-PCR)、Western blot 和免疫荧光技术分别检测目的基因及其蛋白表达。噻唑蓝(MTT)比色法和平板法集落形成实验检测 sTGF-βRⅡ对LoVo 细胞增殖的影响,Annexin V-FITC 和 PI 双染流式细胞术检测 sTGF-βRⅡ对 LoVo 细胞凋亡的影响,Matrigel^(TM)侵袭试剂盒检测 sTGF-βRⅡ对 LoVo 细胞侵袭能力的影响。结果 sTGF-βRⅡ在LoVo 细胞中获得表达。sTGF-βRⅡ可抑制 TGF-β1作用下的 LoVo 细胞增殖、凋亡和体外侵袭能力。结论 sTGF-βRⅡ可拮抗 TGF-β1的信号传导,对大肠癌 LoVo 细胞的侵袭能力具有一定的抑制作用,在大肠癌的治疗中可能有一定的作用。

Objective To investigate the effects of soluble type Ⅱ transforming growth factor （TGF-β） receptor on biological behavior of colorectal carcinoma cell line LoVo. Methods The pcDNA 3.1 （ ＋ ）/sTGF-βR Ⅱ expression vector was constructed by DNA recombinant technique and was introduced into LoVo cells by lipofectactin-mediated DNA transfection. RT-PCR, Western blot and immunofluo- rescent assay were used to determine sTGF-βR Ⅱ expression. The effect of sTGF-βR Ⅱon proliferation of LoVo cells was tested by MTT assay and the clone forming approaches. The effects of sTGF-βR Ⅱ on apop tosis of LoVo cells was determined by flow cytometry with a double-staining method using FITC conjugated annexin V and PI. The migration and invasion were determined by BD BioCoatTM MatrigelTM invasion chambers. Results sTGF-βR Ⅱ mRNA and protein was expressed in LoVo cells, sTGF-βR Ⅱ could inhibit the proliferation, apoptosis and in vitro invasive ability of LoVo cells treated with TGF-Ⅱ. Conclusion sTGF- βR Ⅱ inhibits TGFβ1 signaling in LoVo cells in vitro and attenuates tumor formation.