摘要
近年研究发现维生素K2（VK2）对肝癌具有抑制作用，但目前VK2的抗肝癌的机理尚不明确。VK2对人肝癌细胞株HepG-2细胞增殖的抑制作用及其机制。具体做法如下：体外培养肝癌HepG-2细胞，分别用不同浓度的VK2（0、5、10、20和40μmol／L）处理培养1～3d，采用台盼蓝拒染法测定各时期的各组细胞的活力；收集20μmol／LVK，作用24h和48h后的HepG-2细胞和对照细胞，抽提DNA电泳检测；收集20μmol／LVK，作用48h后的HepG-2细胞和对照细胞，抽提总RNA，半定量RT—PCR检测抗凋亡基因survivin、bcl-2和促凋亡基因bax的mRNA表达水平。各VK2处理组的活细胞数明显低于对照组（P〈0．05）；经VK2处理的细胞其DNA电泳结果呈明显的梯状条带；与对照组相比，VK2处理组细胞的抗凋亡基因survivin和bcl-2的RT—PCR产物电泳的目的条带相对灰度值（目的基因灰度／GAPDH灰度）明显减少（P〈0．05），而促凋亡基因bax则无明显变化（P〉0．05）。VK2通过下调抗凋亡基因survivin和bel－2／bax的桌浊水平诱导肝痛HenG-2细胞凋亡。
It was reported that VK2 could inhibit the proliferation of liver cancer, but at present the mechanism is not known. The inhibitory effects of VK2 on the proliferation of HepG-2 cells and their possible mechanism were studied. The viability of cells treated with various concentration of VK2 （0, 5, 10, 20 and 40μmol/L） from one day to three days by Trypan Blue Exclusion Assay was described. The DNA of cells treated with 0 and 20μmol/L VK2 for 24 and 48 h was extracted and analyzed by eleetrophoresis. Then the levels of bcl-2, survivin and bax transcripts in HepG-2 cells treated with 20μmol/L VK2 for 48 h were determined by semiquantitative reverse transeriptase （RT） -PCR analysis. Results showed that the viability of HepG-2 cells was significantly reduced by VK2 compared with that of the untreated control （p 〈 0. 05 ）. There was DNA ladder in cells with VK2. And the relative densitometric values （ target gene/GAPDH gene） of bel-2 and survivin transcripts in ceils with VK2 were significantly lower than that of the control （p 〈 0. 05 ） , but the relative densitometric value of bax had no significant difference （ p 〉 0. 05 ）. It indicated that VK2 can inhibit the proliferation of HepG-2 cells mediated by apoptosis which is induced by regulating down the levels of survivin and bcl/bax transcripts.
出处
《生物学杂志》
CAS
CSCD
2009年第5期30-33,共4页
Journal of Biology
