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目的:研究蛋白激酶B(AKT)活化诱导骨桥蛋白(OPN)表达在肝细胞肝癌(HCC)骨转移中的作用及其机制。方法:收集40例HCC骨转移灶及其原发灶标本,并另选取22例无转移的HCC组织标本作为对照,行免疫组化染色检测AKT、OPN表达水平;同时用miR-21表达载体转染人HCC细胞系BEL-7402,诱导AKT磷酸化激活,同时用AKT特异性抑制剂(MK-2206)抑制AKT磷酸化激活,分别观察AKT磷酸化水平(p-AKT)与OPN表达间的关系;采用OPN特异性小干扰RNA(OPN-siRNA)沉默BEL-7402细胞OPN表达,并用MTT法检测其对细胞增殖的影响。结果:在发生HCC骨转移灶中,p-AKT和OPN阳性率分别为85.00%(34/40)和77.50%(31/40),明显高于其相应的HCC原发灶组织(p-AKT为60.00%;OPN为55.00%,P<0.05)及无转移的HCC组织(p-AKT为54.55%;OPN为36.36%,P<0.01);BEL-7402细胞转染miR-21诱导p-AKT上调后,OPN表达升高并促进肿瘤细胞增殖加速;反之,MK-2206抑制AKT磷酸化后,BEL-7402细胞OPN表达下降,肿瘤细胞增殖速率降低;采用特异siRNA沉默OPN表达,同样可抑制肿瘤细胞增殖。结论:HCC细胞OPN高表达与AKT信号通路的激活有关,AKT磷酸化水平的升高能诱导OPN表达,进而促进HCC细胞的增殖和转移,提示检测p-AKT与OPN的表达水平对于HCC恶性生物学行为的判断具有潜在的应用价值。
AIM: To investigate the role of AKT activation in inducing osteopontin (OPN) expression in bone metastasis of hepatocellular carcinoma (HCC) and its mechanism. Methods: 40 HCC bone metastases and their primary tumor specimens were collected. Twenty-two non-metastatic HCC tissues were also selected as control. The expression of AKT and OPN was detected by immunohistochemistry. Meanwhile, miR-21 expression vector was used The expression of AKT phosphorylation (AKT) and the expression of OPN in AKT cells were detected by immunofluorescence staining. The phosphorylation of AKT was induced by HIF-2α-Akt staining and the AKT phosphorylation was inhibited by Akt specific inhibitor (MK-2206) ; OPN-siRNA was used to silence OPN expression in BEL-7402 cells, and the effect of OPN on cell proliferation was detected by MTT assay. RESULTS: The positive rates of p-AKT and OPN in bone metastases of HCC were 85.00% (34/40) and 77.50% (31/40), respectively, which were significantly higher than those in the corresponding primary HCC tissues (p-AKT (P <0.05), and the expression of p-AKT was significantly increased in BEL-7402 cells transfected with miR-21 (60.00%; OPN was 55.00% On the contrary, MKN-2206 inhibited AKT phosphorylation, BEL-7402 cells OPN expression decreased, the rate of tumor cell proliferation decreased; using specific siRNA silencing OPN expression, the same can inhibit the tumor Cell Proliferation. CONCLUSION: Overexpression of OPN in HCC cells is related to the activation of AKT signaling pathway. The increase of phosphorylation of AKT can induce the expression of OPN, and then promote the proliferation and metastasis of HCC. It is suggested that the expression of p-AKT and OPN may play an important role in HCC malignant biology Judgment of behavior has potential application value.