BCL6B对人结直肠癌LoVo细胞增殖和迁移的影响及机制

摘要

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 BCL6B on proliferation and migration of LoVo cells for further explo-ring the underlying mechanism .METHODS:The endogenous expression of BCL 6B in the FHC and LoVo cells was detec-ted 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 BCL 6B in the LoVo cells.The mRNA and protein levels of BCL6B, cyclin D1 and matrix metalloproteinase-9 ( MMP-9) were determined by RT-PCR and Western blot , re-spectively.The level of phosphorylated protein kinase B (p-AKT) was detected by Western blot.RESULTS:BCL6B ex-pression was notably repressed in the LoVo cells as compared with the FHC cells , which were significantly increased by transfection with pcDNA3.1-BCL6B.The abilities of proliferation and migration of the LoVo cells at 72 h were inhibited by 28.33%(P<0.01) and 36.11%(P<0.05) in BCL6B group.The mRNA levels of cyclin D1 and MMP-9 in the cells of BCL6B 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:BCL6B inhibits proliferation and migration of the LoVo cells , and the PI3K/AKT signaling pathway is in-volved in this process .%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pcDNA3．1－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pcD－NA3．1－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mRNA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。