5种致癌相关基因在乙型肝炎病毒感染进程中的差异表达

Differential expression of five kinds of carcinoma-related genes in hepatits B virus infection progression

目的探讨致癌相关基因在慢性乙型肝炎(chronic hepetitis B,CHB)患者、乙型肝炎病毒(hepatitis B virus,HBV)携带者、HBV相关肝衰竭患者及健康人群外周血白细胞中的差异表达。方法收集2019年2月至2019年9月云南省第二人民医院收治的51例CHB患者(记为CHB组)、51例HBV携带者(记为ASC组)、15例HBV相关肝衰竭患者(记为LF组)以及51例健康体检者(HC组)为研究对象。采集所有研究对象的外周血,提取外周血白细胞内的总RNA。采用定量反转录聚合酶链式反应(quantitative reverse transcriptase-mediated polymerase chain reaction,qRT-PCR)法检测DNAJC6、GSE、HIPK3、MBOAT2、UBXN7线状RNA(liner RNA)、环状RNA(circular RNA,circRNA)的相对表达水平,并评估其环线比在各组间的变化。结果与HC组相比,ASC组Lin-DNAJC6(3.59±1.47 vs 2.36±1.10)、Lin-HIPK3(1.79±0.71 vs 1.33±0.65)、Lin-UBXN7(8.68±1.96 vs 3.36±0.86)、Circ-GSE1(2.71±1.47 vs 1.83±1.09)、Circ-UBXN7(2.88±1.37 vs 2.03±1.19)相对表达量显著升高,Lin-GSE1(1.65±0.94 vs 2.04±0.79)、Circ-HIPK3(1.93±1.09 vs 3.85±1.23)相对表达量显著降低(P均<0.05);CHB组Lin-DNAJC6(6.73±1.48 vs 2.36±1.10)、Lin-HIPK3(7.24±0.85 vs 1.33±0.65)、Lin-MBOAT2(6.58±0.72 vs 1.35±0.70)、Circ-DNAJC6(5.26±1.86 vs 3.41±1.75)、Circ-GSE1(2.61±1.68 vs 1.83±1.09)、Circ-MBOAT2(5.50±1.82 vs 3.22±1.52)、Circ-UBXN7(4.61±1.36 vs 2.03±1.19)相对表达量显著升高,Lin-UBXN7(2.47±0.83 vs 3.36±0.86)相对表达量显著降低(P均<0.05);LF组Lin-DNAJC6(5.10±2.49 vs 2.36±1.10)、Lin-HIPK3(2.47±1.22 vs 1.33±0.65)、Circ-GSE1(2.98±2.12 vs1.83±1.09)相对表达量显著升高,Circ-HIPK3(2.29±1.40 vs 3.85±1.23)相对表达量显著降低。各组DNAJC6、GSE1、HIPK3、MBOAT2、UBXN7的RNA环线比存在不同程度的差异。ASC组GSE1的环线比(中位数:1.77 vs 0.93)显著高于HC组,DNAJC6(中位数:1.10 vs 1.52)、HIPK3(中位数:1.05 vs 2.95)、UBXN7(中位数:0.31 vs 0.56)的环线比显著低于HC组(P均<0.05);CHB组GSE1(中位数1.43 vs 0.93)、UBXN7(中位数:1.82 vs 0.56)的环线比显著高于HC组(P均<0.05),DNAJC6(中位数:0.78 vs 1.52)、HIPK3(中位数:0.50 vs 2.95)、MBOAT2(中位数:0.82 vs 2.14)的环线比显著低于HC组(P均<0.05);LF组DNAJC6(中位数:0.65 vs 1.52)环线比显著低于对照组(P<0.05)。结论DNAJC6、GSE1、HIPK3、MBOAT2、UBXN7线状RNA和circRNA在HBV感染肝病患者外周血白细胞中差异表达,提示circRNA可能与HBV感染相关肝病的进展密切相关。

Objective To investigate the differential expression of carcinoma-related genes in peripheral blood leukocytes of patients with chronic hepetitis B(CHB),hepatitis B virus(HBV)carriers,patients with HBV-related liver failure and healthy controls.Methods Patients with CHB(CHB group,51 cases),HBV carriers(ASC group,51 cases),patients with HBVrelated liver failure(LF group,15 cases)and healthy controls(HC group,51 cases)in the Second People’s Hospital of Yunnan Province from February 2019 to September 2019 were enrolled.Peripheral blood was collected from all study subjects and total RNA was extracted from peripheral blood leukocytes.The relative expression levels of DNAJC6,GSE1,HIPK3,MBOAT2,UBXN7 linear and circular RNA were detected by quantitative reverse transcriptasemediated polymerase chain reaction(qRT-PCR),and the changes of circRNA/linear RNA expression ratio among groups were evaluated.Results Compared with healthy controls,the relative expression levels of Lin-DNAJC6(3.59±1.47 vs 2.36±1.10),Lin-HIPK3(1.79±0.71 vs 1.33±0.65),Lin-UBXN7(8.68±1.96 vs 3.36±0.86),Circ-GSE1(2.71±1.47 vs 1.83±1.09)and Circ-UBXN7(2.88±1.37 vs 2.03±1.19)were significant higher in ASC group,while the relative expression levels of Lin-GSE1(1.65±0.94 vs 2.04±0.79)and Circ-HIPK3(1.93±1.09 vs 3.85±1.23)were significantly lower(all P<0.05).Compared with healthy controls,the relative expression levels of Lin-DNAJC6(6.73±1.48 vs 2.36±1.10),Lin-HIPK3(7.24±0.85 vs 1.33±0.65),Lin-MBOAT2(6.58±0.72 vs 1.35±0.70),Circ-DNAJC6(5.26±1.86 vs 3.41±1.75),Circ-GSE1(2.61±1.68 vs 1.83±1.09),Circ-MBOAT2(5.50±1.82 vs 3.22±1.52)and Circ-UBXN7(4.61±1.36 vs 2.03±1.19)in CHB group were significantly higher,while the relative expression level of Lin-UBXN7(2.47±0.83 vs 3.36±0.86)was significantly lower(all P<0.05).Compared with healthy controls,the relative expression levels of Lin-DNAJC6(5.10±2.49 vs 2.36±1.10),Lin-HIPK3(2.47±1.22 vs 1.33±0.65)and Circ-GSE1(2.98±2.12 vs 1.83±1.09)were significantly higher in LF group,while the relative expression levels of Circ-HIPK3(2.29±1.40 vs 3.85±1.23)was significantly lower(all P<0.05).Meanwhile,the circRNA/linear RNA expression ratio of DNAJC6,GSE1,HIPK3,MBOAT2,UBXN7 were different in each group.The circRNA/linear RNA expression ratio of GSE1(median:1.77 vs 0.93)in ASC group were significantly higher than HC group(P<0.05).The circRNA/linear RNA expression ratio of DNAJC6(median:1.10 vs 1.52),HIPK3(median:1.05 vs 2.95)and UBXN7(median:0.31 vs 0.56)in ASC group were significantly lower than HC group(all P<0.05).The circRNA/linear RNA expression ratio of GSE1(median:1.43 vs 0.93)and UBXN7(median:1.82 vs 0.56)in patients with CHB were significantly higher than those of healthy controls(all P<0.05).The circRNA/linear RNA expression ratio of DNAJC6(median:0.78 vs 1.52),HIPK3(madian:0.50 vs 2.95)and MBOAT2(median:0.82 vs 2.14)in patients with CHB were significantly lower than those of healthy controls(all P<0.05).The circRNA/linear RNA expression ratio of DNAJC6(median:0.65 vs 1.52)of patients in LF group were significantly lower than those of healthy controls(P<0.05).Conclusions DNAJC6,GSE1,HIPK3,MBOAT2,UBXN7 linear RNA and circRNA were differentially expressed in peripheral blood leukocytes of patients with HBV-infected liver disease,suggesting that circRNA may be closely related to the progression of liver disease associated with HBV infection.