乙型肝炎病毒对SPINK1表达的影响

The effect of hepatitis B virus on the expression of SPINK1

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 explore the effect of hepatitis B virus（ HBV） on the expression of SPINK1 and its regulatory mechanism. Methods RT- PCR and Western blot were used to measure the mRNA and protein expression of SPINK1 in Hep G2 and Hep G2. 2. 15 cells; serum SPINK1 levels in patients with HBV infection and in healthy controls were measured by enzyme-linked immunosorbent assay（ ELISA）,and the differences of serum SPINK1 levels among patients with chronic hepatitis B（ CHB）,liver cirrhosis（ LC） and hepatocellular carcinoma（ HCC） were analyzed; Hep G2 cells were co- transfected with SPINK1 promoter containing the luciferase gene and HBV infectious clone p HBV1. 3,luciferase activity,mRNA and protein expression of SPINK1 were measured. Results Expression of SPINK1 was higher in Hep G2. 2. 15 cells than in Hep G2 cells; serum SPINK1 level was much higher in HBV patients than in the healthy controls,and the difference was statistically significant（ P〈0. 05）; SPINK1 was detected at higher levels in patients with LC and HCC than those with and CHB; HBV can enhance SPINK1 gene promoter activity and up- regulate the SPINK1 expression of mRNA and protein. Conclusion HBV up- regulates the expression of SPINK1 via enhancing SPINK1 gene promoter activity,and the serum levels of SPINK1 are associated with the progression of HBV infection.