论文部分内容阅读
目的探讨乙型肝炎病毒(HBV)对丝氨酸转肽酶抑制剂Kazal 1型(SPINK1)表达的影响及其调节机制。方法采用逆转录PCR(RT-PCR)和免疫印迹法(Western blot),检测Hep G2和Hep G2.2.15细胞中SPINK1 mRNA和蛋白的表达;通过酶联免疫吸附试验(ELISA)检测HBV患者和健康对照者SPINK1血清学水平,比较分析慢性乙型肝炎(CHB)、肝纤维化(LC)和肝癌患者(HCC)血清SPINK1含量的差异;将HBV感染性克隆p HBV1.3与SPINK1基因启动子共转染Hep G2细胞,测定荧光素酶的活性,检测SPINK1 mRNA和蛋白的表达。结果 Hep G2.2.15细胞中SPINK1mRNA和蛋白的表达水平较Hep G2高;SPINK1在HBV患者血清学水平显著升高,差异有统计学意义(P<0.05);SPINK1在肝纤维化患者和肝癌患者明显高于慢性肝炎患者;HBV能够激活SPINK1基因启动子的活性,并在mRNA和蛋白水平上调SPINK1的表达。结论 HBV通过上调SPINK1启动子的活性调节SPINK1的表达,其血清学水平与HBV疾病进程相关。
Objective To investigate the effect of hepatitis B virus (HBV) on the expression of serine peptidase inhibitor Kazal 1 (SPINK1) and its regulatory mechanism. Methods The expression of SPINK1 mRNA and protein in Hep G2 and Hep G2.2.15 cells was detected by RT-PCR and Western blot. The levels of SPINK1 mRNA and protein in Hep G2 and Hep G2.2.15 cells were detected by enzyme linked immunosorbent assay (ELISA) Control group SPINK1 serological level, comparative analysis of chronic hepatitis B (CHB), liver fibrosis (LC) and liver cancer patients (HCC) serum SPINK1 content differences; the HBV infectious clone p HBV1.3 and SPINK1 gene promoter Hep G2 cells were transfected, the luciferase activity was measured, and the expression of SPINK1 mRNA and protein was detected. Results The expression level of SPINK1 mRNA and protein in Hep G2.2.15 cells was higher than that in Hep G2 cells. The serological level of SPINK1 in HBV patients was significantly higher than that in Hep G2 cells (P <0.05); SPINK1 was significantly higher in patients with liver fibrosis and liver cancer Higher than chronic hepatitis patients; HBV can activate the activity of SPINK1 gene promoter and up-regulate the expression of SPINK1 at mRNA and protein level. Conclusion HBV regulates the expression of SPINK1 by up-regulating the activity of SPINK1 promoter. The serological level of HBV is related to the progression of HBV diseases.