载脂蛋白 AI 和 AIV 在肝细胞癌中的表达分析：基于蛋白质的肝细胞癌相关研究。
Expression analysis of apolipoproteins AI & AIV in hepatocellular carcinoma: A protein-based hepatocellular carcinoma-associated study.
机构信息
Department of Medicine, Maulana Azad Medical College, University of Delhi, New Delhi, India.
Department of Biochemistry, University College of Medical Sciences, Delhi, India.
出版信息
Indian J Med Res. 2018 Apr;147(4):361-368. doi: 10.4103/ijmr.IJMR_1358_16.
BACKGROUND & OBJECTIVES: Hepatocellular carcinoma (HCC) is one of the leading causes of cancer mortality. The objective of this study was to find out the differential expression of apolipoproteins (ApoAI and ApoAIV) in HCC and cases of liver cirrhosis and chronic hepatitis (controls) without HCC and to compare ApoAI and ApoAIV expression with alpha-foetoprotein (AFP), the conventional marker in HCC.
METHODS
Fifty patients with HCC and 50 controls comprising patients with liver cirrhosis (n=25) and chronic hepatitis (n=25) without HCC were included in this study. Total proteins were precipitated using acetone precipitation method followed by albumin and IgG depletion of precipitated protein using depletion kit. Proteins were separated by sodium dodecyl sulphate-polyacrylamide gel electrophoresis. The expression changes of ApoAI and ApoAIV were confirmed by western blotting using specific primary and secondary polyclonal antibodies followed by densitometric protein semi-quantitative estimation. ApoAI, ApoAIV and AFP were measured in the plasma samples by ELISA method.
RESULTS
Semi-quantitative densitometric image analysis of the western blot images and the comparison between HCC patients with those without HCC (control) revealed differential expression of ApoAI and ApoAIV. Levels of ApoAI were significantly higher in patients with HCC compared to controls without HCC (0.279±0.216 vs 0.171±0.091 and 0.199±0.014; P <0.001). Levels of ApoAIV were significantly lower in patients of HCC compared to controls without HCC (0.119±0.061 vs 0.208±0.07 and 0.171±0.16; P <0.01). ELISA assays of apolipoproteins (ApoAI and ApoAIV) revealed similar results of expression of ApoAI and ApoAIV as detected in western blotting densitometric image analysis.
INTERPRETATION & CONCLUSIONS: Increased expression of ApoAI and decreased expression of ApoAIV in HCC patients compared to controls without HCC revealed the abnormalities in HCC. These molecules need to be studied further for their use as potential biomarkers in the future diagnostic tools along with other conventional biomarkers for screening of HCC cases. It needs further analysis in higher number of patient population.
背景与目的
肝细胞癌（HCC）是癌症死亡的主要原因之一。本研究旨在确定载脂蛋白（ApoAI 和 ApoAIV）在 HCC 以及无 HCC 的肝硬化和慢性肝炎（对照）病例中的差异表达，并将 ApoAI 和 ApoAIV 的表达与甲胎蛋白（AFP）进行比较，后者是 HCC 的常规标志物。
方法
本研究纳入了 50 例 HCC 患者和 50 例对照，包括 25 例肝硬化患者和 25 例无 HCC 的慢性肝炎患者。使用丙酮沉淀法沉淀总蛋白，然后使用沉淀试剂盒去除沉淀蛋白中的白蛋白和 IgG。用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳分离蛋白质。使用特异性的初级和次级多克隆抗体通过 Western 印迹法确认 ApoAI 和 ApoAIV 的表达变化，然后进行密度计蛋白半定量估计。通过 ELISA 法测量血浆样本中的 ApoAI、ApoAIV 和 AFP。
结果
对 Western 印迹图像的半定量密度图像分析以及 HCC 患者与无 HCC（对照）患者的比较显示，ApoAI 和 ApoAIV 存在差异表达。与无 HCC 的对照组相比，HCC 患者的 ApoAI 水平显著升高（0.279±0.216 vs 0.171±0.091 和 0.199±0.014；P <0.001）。与无 HCC 的对照组相比，HCC 患者的 ApoAIV 水平显著降低（0.119±0.061 vs 0.208±0.07 和 0.171±0.16；P <0.01）。载脂蛋白（ApoAI 和 ApoAIV）的 ELISA 检测结果显示，ApoAI 和 ApoAIV 的表达与 Western 印迹密度图像分析结果相似。
解释与结论
与无 HCC 的对照组相比，HCC 患者中 ApoAI 的表达增加和 ApoAIV 的表达减少表明 HCC 存在异常。这些分子需要进一步研究，以作为未来 HCC 筛查工具中与其他常规生物标志物一起使用的潜在生物标志物。需要在更多的患者人群中进行进一步分析。
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