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目的观察肝癌组织中维生素K缺乏或拮抗剂-Ⅱ诱导的蛋白质(protein induced by vitamin K absence or antagonist-Ⅱ,PIVKA-Ⅱ)表达情况及维生素K_2(vitamin K_2,VitK_2)对肝癌细胞中PIVKA-Ⅱ的影响,探讨PIVKA-Ⅱ与肝癌的关系及可能机制。方法选择浙江省肿瘤医院、浙江省中山医院2012年1月—2015年12月100对手术切除的原发性肝癌组织及癌旁组织标本,采用ABC免疫组织化学法测定组织中PIVKA-Ⅱ的阳性表达情况,采用化学发光免疫分析法测定标本中PIVKA-Ⅱ水平;将HepG-2细胞和浓度为20μM的VitK_2共同培养,酶联免疫吸附(ELISA)法测定VitK_2对HepG-2肝癌细胞PIVKA-Ⅱ表达的影响,MTT法测定VitK_2对HepG-2肝癌细胞生长的影响,Transwell法测定VitK_2对HepG-2肝癌细胞侵袭能力的影响。结果肝癌组织中PIVKA-Ⅱ阳性率(74.0%)高于癌旁组织中PIVKA-Ⅱ阳性表达率(25.0%,P<0.05);肝癌组织中PIVKA-Ⅱ水平(3 786.23±143.24)m AU/g高于癌旁组织中PIVKA-Ⅱ水平(167.34±21.54)m AU/g,P<0.05。对照组肝癌细胞PIVKA-Ⅱ水平为(3.43±0.04)ng/(ml·10~6细胞),VitK_2组肝癌细胞PIVKA-Ⅱ水平为(2.57±0.02)ng/(ml·10~6细胞),VitK_2组肝癌细胞PIVKA-Ⅱ水平明显低于对照组(P<0.05)。VitK_2与HepG-2肝癌细胞共同培养,第1天、第2天、第3天、第4天、第5天、第6天时VitK_2对HepG-2细胞的抑制率分别为9.2%、16.5%、26.7%、34.8%、41.2%、46.7%。VitK_2与HepG-2肝癌细胞培养24 h后,VitK_2对HepG-2肝癌细胞侵袭的抑制率为37.2%。结论肝癌组织中PIVKA-Ⅱ呈高表达,VitK_2能够降低肝癌细胞中PIVKA-Ⅱ水平、抑制肝癌细胞生长、降低肝癌细胞侵袭能力。
Objective To observe the expression of PIVK-Ⅱ or PIVKA-Ⅱ induced by vitamin K deficiency or antagonist-Ⅱ and the effect of vitamin K_2 (VitK_2) on the expression of PIVKA-Ⅱ , To explore the relationship between PIVKA-Ⅱ and hepatocellular carcinoma and its possible mechanism. Methods 100 cases of primary hepatocellular carcinoma and adjacent non-cancerous tissue specimens from Zhejiang Cancer Hospital and Zhongshan Hospital of Zhejiang Province from January 2012 to December 2015 were selected and the positive expression of PIVKA-Ⅱ in tissues was detected by ABC immunohistochemical method The levels of PIVKA-Ⅱ in HepG-2 hepatoma cells were determined by enzyme-linked immunosorbent assay (ELISA). The expression of PIVKA-Ⅱ was detected by chemiluminescence immunoassay. HepG-2 cells were co-cultured with VitK 2 at a concentration of 20 μM. The effect of VitK_2 on the growth of HepG-2 hepatoma cells was determined by MTT assay. The effect of VitK_2 on the invasion ability of HepG-2 hepatocellular carcinoma cells was determined by Transwell assay. Results The positive rate of PIVKA-Ⅱ (74.0%) in HCC tissues was higher than that in PITA tissues (25.0%, P <0.05). The PIVKA-Ⅱ level in HCC tissues (3 786.23 ± 143.24) m AU / g was significantly higher than PIVKA-Ⅱ (167.34 ± 21.54) m AU / g, P <0.05. The level of PIVKA-Ⅱ was (3.43 ± 0.04) ng / (ml · 10 ~ 6) in control group, PIVKA-Ⅱ was (2.57 ± 0.02) ng / (· 10 · 6) in VitK_2 group, The level of PIVKA-Ⅱ in VitK_2 group was significantly lower than that in control group (P <0.05). VitK_2 and HepG-2 cells were co-cultured. The inhibitory rates of VitK_2 on HepG-2 cells were 9.2%, 16.5% on day 1, day 2, day 3, day 4, day 5 and day 6, 26.7%, 34.8%, 41.2%, 46.7%. VitK_2 and HepG-2 hepatoma cells cultured for 24 h, VitK 2 HepG-2 hepatoma cell invasion inhibition rate of 37.2%. Conclusions PIVKA-Ⅱ is highly expressed in hepatocellular carcinoma. VitK_2 can reduce the level of PIVKA-Ⅱ in hepatoma cells, inhibit the growth of hepatoma cells and decrease the invasiveness of hepatocellular carcinoma cells.