骨形态发生蛋白5对肝癌细胞生物学行为的影响及机制  

Effect of BMP5 on biological behaviors of liver cancer cells and its mechanism

作　　者：马东玥 臧艳[1] 任俏慧 朱欣悦 王莲子 吕伟 姚骏骁 周心怡 余莉 李涛[1] MA Dongyue;ZANG Yan;REN Qiaohui;ZHU Xinyue;WANG Lianzi;LYU Wei;YAO Junxiao;ZHOU Xinyi;YU Li;LI Tao(Department of Clinical Laboratory,The First Affiliated Hospital of Anhui Medical University,Hefei 230022,China;不详)

机构地区：[1]安徽医科大学第一附属医院检验科,合肥230022 [2]病原生物学安徽省重点实验室

出　　处：《山东医药》2024年第10期1-5,共5页Shandong Medical Journal

基　　金：安徽省转化医学研究院科研基金项目(2023zhyx-C24);安徽医科大学开放研究基金重点项目(KFJJ-2021-04);安徽省学术和技术带头人及后备人选科研活动经费资助项目(2022H291)。

摘　　要：目的 探究骨形态发生蛋白5(BMP5)对人肝癌细胞生物学行为的影响及机制。方法 将正常人肝细胞系L02与人肝癌细胞系Hep3B、Huh7置于37℃、5%CO_(2)细胞培养箱中培养,采用RT-qPCR法检测正常人肝细胞系L02与人肝癌细胞系Hep3B、Huh7中BMP5 mRNA表达。将肝癌细胞系Hep3B、Huh7分为对照(NC)组及低、中、高剂量组,低、中、高剂量组分别用1、10、100 ng/mL浓度的重组人骨形态发生蛋白5(rh-BMP5)处理细胞,通过CCK-8法检测细胞增殖,Annexin-V/PI流式细胞术检测细胞凋亡,划痕实验检测细胞迁移,Western blotting法检测增殖细胞核抗原PCNA和SMAD通路关键蛋白及其磷酸化(SMAD3、p-SMAD3)表达。结果 与正常肝细胞系L02相比,BMP5在肝癌细胞系Hep3B、Huh7中表达升高(P均<0.05)。与NC组相比,中、高剂量组rh-BMP5可促进Hep3B、Huh7细胞增殖和PCNA表达上调(P均<0.05),高剂量组100 ng/mL的rh-BMP5处理肝癌细胞24及48 h细胞凋亡率降低、迁移率增高(P均<0.05)。高剂量组与NC组相比,p-SMAD3表达升高(P<0.05)。结论 BMP5在肝癌细胞系中高表达,可能通过增强SMAD3活化促进肝癌细胞Hep3B、Huh7的增殖、迁移并抑制凋亡。Objective To investigate the effect and mechanism of bone morphogenetic protein 5(BMP5) on the biological behaviors of human liver cancer cells.Methods Normal human liver cell line L02 and human liver cancer cell lines Hep3B and Huh7 were cultured in the incubator at 37℃ with 5% CO_2.The expression of BMP5 mRNA in these cells was detected by real time-quantitive PCR(RT-qPCR).Human liver cancer cells Hep3B and Huh7 were divided into the control group(NC group),low-dose group,medium-dose group and high-dose group.Cells in the low-,medium-and high-dose groups were treated with recombinant human bone morphogenetic protein 5(rh-BMP5) at concentrations of 1,10 and 100 ng/mL,respectively.Cell proliferation was detected by CCK-8 assay and apoptosis was detected by AnnexinV/PI flow cytometry.Scratch assay was used to detect cell migration.Western blotting was performed to detect the expression of proliferating cell nuclear antigen(PCNA) and key molecules in SMAD pathway(SMAD3) and its phosphorylation(p-SMAD3).Results The expression levels of BMP5 in Hep3B and Huh7 cells significantly increased in comparison with that in L02 cells(both P<0.05).Compared with NC group,rh-BMP5 promoted the proliferation of Hep3B and Huh7cells and up-regulated the expression of PCNA in the medium-dose and high-dose groups(all P<0.05).After the highdose(100 ng/mL) rh-BMP5 treatment,Hep3B and Huh7 cells showed decreased apoptosis rate and increased migration rate at 24 h and 48 h in comparison with those of the NC group(both P<0.05).The high-dose rh-BMP5 group had higher p-SMAD3 level than the NC group(P<0.05).Conclusions The expression of BMP5 increases in liver cancer cell lines.BMP5 promotes proliferation and migration and inhibits apoptosis of Hep3B and Huh7 cells,which may be mediated by SMAD3 phosphorylation,and thus facilitates tumor progression.

关 键 词：肝癌 骨形态发生蛋白5 细胞增殖 SMAD同源物3 

分 类 号：R735.7[医药卫生—肿瘤]