BCL6B对人结直肠癌LoVo细胞增殖和迁移的影响及机制

Effects of BCL6B on proliferation and migration of human colorectal carcinoma LoVo cells and its potential mechanism

目的:研究B细胞白血病/淋巴瘤6B(BCL6B)在人正常肠上皮细胞FHC及结直肠癌细胞LoVo中的表达水平及BCL6B对LoVo细胞增殖和迁移的影响,并探讨其相关分子机制。方法:采用RT-PCR及Western blot检测FHC细胞和LoVo细胞中BCL6B的内源性表达;用脂质体法将重组质粒pc DNA3.1-BCL6B转入LoVo细胞,运用MTT、集落形成、细胞划痕及Transwell小室实验检测BCL6B对LoVo细胞增殖和迁移的影响,采用RT-PCR及Western blot检测细胞周期蛋白D1(cyclin D1)和基质金属蛋白酶9(MMP-9)的表达,Western blot检测蛋白激酶B(AKT)的磷酸化水平。结果:与正常肠上皮细胞FHC相比,BCL6B在LoVo细胞中呈明显低表达;转染pc DNA3.1-BCL6B后的LoVo细胞内BCL6B水平显著增高。过表达BCL6B的实验组细胞72 h的增殖活性及划痕愈合力分别较对照组降低28.33%(P<0.01)和36.11%(P<0.05)。实验组细胞cyclin D1和MMP-9的m RNA水平分别降低39.90%(P<0.01)和77.36%(P<0.05),同时cyclin D1、MMP-9和磷酸化蛋白激酶B(p-AKT)的蛋白水平分别降低44.00%(P<0.05)、47.06%(P<0.01)和32.88%(P<0.05)。结论:BCL6B可抑制结直肠癌LoVo细胞的增殖和迁移,其机制可能涉及PI3K/AKT信号通路的抑制。

AIM：To detect the endogenous expression of B-cell leukemia/lymphoma 6 member B（BCL6B）in FHC and LoVo cells,and to investigate the effects of BCL6 B on proliferation and migration of LoVo cells for further exploring the underlying mechanism. METHODS：The endogenous expression of BCL6 B in the FHC and LoVo cells was detected by RT-PCR and Western blot. The methods of MTT assay,colony formation assay,wound healing assay and Transwell chamber experiment were employed to examine the biological functions of BCL6 B in the LoVo cells. The m RNA and protein levels of BCL6 B,cyclin D1 and matrix metalloproteinase-9（MMP-9）were determined by RT-PCR and Western blot,respectively. The level of phosphorylated protein kinase B（p-AKT）was detected by Western blot. RESULTS：BCL6 B expression was notably repressed in the LoVo cells as compared with the FHC cells,which were significantly increased by transfection with pc DNA3. 1-BCL6 B. The abilities of proliferation and migration of the LoVo cells at 72 h were inhibited by28. 33%（P 〈 0. 01）and 36. 11%（P 〈 0. 05）in BCL6 B group. The m RNA levels of cyclin D1 and MMP-9 in the cells of BCL6 B group were decreased by 39. 90%（P 〈 0. 01）and 77. 36%（P 〈 0. 05）,and the protein levels of cyclin D1,MMP-9 and p-AKT were reduced by 44. 00%（P 〈 0. 05）,47. 06%（P 〈 0. 01）and 32. 88%（P 〈 0. 05）,respectively.CONCLUSION：BCL6 B inhibits proliferation and migration of the LoVo cells,and the PI3 K/AKT signaling pathway is involved in this process.