目的：探究肝细胞癌中分泌性富含半胱氨酸的酸性蛋白（ secreted protien acidic and rich in cysteine ， SPARC）表达及其与糖酵解作用的相关调节机制。方法选取肝癌细胞HepG2、Hep3B、Huh7，分别稳定转染SPARC质粒和SPARC siRNA，应用比色法检测2种转染的细胞中糖酵解关键酶己糖激酶（ HK）和乳酸脱氢酶-A（ LDHA）的活性变化。结果转染SPARC质粒的3种肝癌细胞中HK-Ⅱ、LDHA的活性较对照组明显下降，3种细胞中HK-Ⅱ相对吸光度分别降低至（57．45±5．05）％，（56．1±5．70）％，（49．73±7．06）％；LDHA相对吸光度分别降低至（50．53±8．07）％，（54．38±2．08）％，（42．34±3．44）％；差异均有统计学意义（P＜0．05）；反之，siRNA抑制SPARC表达的肝癌细胞中， HK-Ⅱ、LDHA活性较对照组明显上调，3种细胞中 HK-Ⅱ相对吸光度分别上调至（174．20±8．41）％，（166．69±9．71）％，（152．84±3．39）％；LDHA 的相对吸光度分别上调至（176．65±6．85）％；（163．24±6．44）％；（151．25±6．75）％，差异具有统计学意义（P＜0．05）。结论肝细胞癌中，SPARC通过负性调控糖酵解作用的关键酶抑制糖酵解。%Objective To investigate the expression of SPARC in hepatocellular carcinoma and its correlation with regu-lating mechanism of glycolysis.Methods HepG2,Hep3B,Huh7 human hepatocellular carcinoma cells were stably transfected with vector containing wild type SPARC or siRNA SPARC ,and the activity of HK-Ⅱand LDHA were detected by colourimetric method.Results The activity of HK-Ⅱand LDHA significantly decreased in the 3 kinds of hepatocellular carcinoma cells trans-fected with SPARC,the HK relatively absorbance in the 3 kinds of cells were down to (57.45 ±5.05)%,(56.1 ±5.70)%and (49.73 ±7.06)%,respectively;the LDHA relatively absorbance were down to (50.53 ±8.07)%,(54.38 ±2.08)% and (42.34 ±3.44)%,there had statistical difference (P<0.05);and the reverse results were observed in the 3 kinds of hepatocel-lular carcinoma cells when SPARC was inhibited by siRNA ,the HK relatively absorbance roze to (174.20 ±8.41)%;(166.69 ± 9.71)%and(152.84 ±3.39)%;the LDHA relatively absorbance roze to (176.65 ±6.85)%;(163.24 ±6.44)% and (151.25 ±6.75)%,there had statistical difference(P<0.05).Conclusion SPARC negatively regulates glucose metabolism by inhibiting the activity of HK-Ⅱand LDHA in hepatocellular carcinoma .
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