己糖激酶2在肝癌转移中的调控作用

The role of hexokinase 2 in the metastasis of hepatocellular carcinoma cells

目的探讨己糖激酶2（HK2）在肝癌中的表达及其在肝癌转移中的调控作川。方法采用免疫组化法分析73例肝癌及癌旁组织中HK2的表达。小十扰RNA（siRNA）干扰HK2的表达后，采用细胞划痕实验检测肝癌细胞的运动能力，Transwell实验检测肝癌细胞的侵袭能力，实时荧光定量聚合酶链式反应和Westernblot检测肝癌细胞中调控上皮间质转化（EMT）关键分子E-cadherin、ZO-1、N—cadherin和vimentin的表达．分析HK2在肝癌细胞EMT中的调控作用。结果免疫组化结果显示．肝癌及癌旁组织中HK2的表达水平分别为5．39±3．40和2．16±1．55，差异有统计学意义（P〈0．05）。siRNA干扰HK2的表达后，肝癌细胞的相对迁移距离由1．00±0．54下降至0．56±0．09，差异有统计学意义（P〈0．05）；细胞侵袭数南（345±42）个下降至（215±34）个，差异有统计学意义（P〈0．05）；肝癌上皮细胞标志物E-eadherin和ZO-1的表达上调，而间质细胞标志物N-cadhefin和vimentin的表达下调。结论HK2在肝癌细胞中的表达升高，通过促进细胞EMT过程而促进肝癌转移。

Objective To investigate the regulatory role of HK2 in the metastasis of hepatocellular carcinoma （HCC）. Methods The protein expressions of HK2 in 73 HCC tumor tissues and paired adjacent non-tumor tissues were evaluated by using immunohistoehemical analysis. The scratch wound healing assay and Transwell assay had been used to analyze the migration and invasion of HCC cells with HK2 knockdown. Expressions of epithelial-to-mesenchymal transition （EMT）markers, such as E-cadherin, ZO-l, N-cadherin and vimentin,in HCC cells with HK2 knockdown were determined by qRT-PCR and Western blot analysis. Results The expression levels of HK2 in tumor tissues and adjacent non-tumor tissues were 5.39±3.40 and 2.16± 1.55, respectively. The protein expression of HK2 was significantly higher in tumor tissues compared with adjacent non-tumor tissues of HCC （P〈0.05）. Knockdown of HK2 in HCC cells decreased the cell motility from 1.00±0.54 to 0.56±0.09 （P〈 0.05 ）, andknuckdown of HK2 in HCC cells decreased the number of invaded cells form 345±42 to 215±34 （P〈0.05）. The expression of epithelial markers ZO-1 and E-cadherin were up-regulated, while mesenchymal markers vimentin and N-cadherin were down-regulated in HCC ceils when HK2 was knockeddown. Conclusions HK2 is up-regulated in HCC and promotes cell motility by stimulating EMT.