基因芯片技术检测肝细胞癌前C区/BCP区基因突变

Study on detection of HBV precore region and basic core promoter region mutations in HCC by using Gene-chips technique

目的应用基因芯片技术探讨HBV慢性感染并发肝细胞癌(HCC)前C区/BCP区基因突变的临床意义。方法应用基因芯片杂交技术检测46例HBV慢性感染并发肝细胞癌前C区A1896、A1899及BCP区nt1762、nt1764四位点突变,比较各位点突变的发生率;据乙肝血清学标志熏将研究对象分为HBeAg阳性组、HBeAg阴性组两组,比较前C区/BCP区变异与HBeAg分泌障碍的关系,分析前C区/BCP区基因突变与血清HBVDNA定量的关系。结果①用基因芯片法测定46例患者,阳性率91.3%(42/46),A1896突变率52.4%,A1899突变率19%熏nt1762/nt1764联合突变率66.7%。②HBeAg阴性组与HBeAg阳性组比较,A1896突变率、nt1762nt1764联合突变率及多位点突变率明显为高,P<0.05。前C区/BCP区变异组与非变异组比较,HBVDNA定量无显著性差异。结论应用基因芯片法可一次同时检测乙型肝炎病毒多个突变位点熏HBV持续慢性感染并发的HCC前C区/BCP区变异的发生率较高,该变异与HBeAg分泌障碍有关,但与HBVDNA复制水平并无明显相关性。

Objective To investigate the clinical significance of HBV precore region and basic core region mutations in hepatocellular carcinoma using Gene-chips technique.Methods Four spots muta-tions of A1896and A1899in precore region and nt1762and nt1764in the basic core promoter region in46cases were detected by Gene chips.Each spot mutation rate was compared.According to HBV marks,all the cases were divided into HBeAg positive group and HBeAg negative group and precore,and BCP variants rate of each group was compared.HBV DNA quantity in each mutant group was compared to the non-mutant group in the meantime.Results①Gene-chips technique can detect different special variant sites of HBV at one time.The positive rate was91.3%(42/46).Mutation rate of A1896was52.4%and mutation rate of A1899was19%.For nt1762and nt1764double variants rate was66.7%.②The rate of A1896and nt1762nt1764double mutations was higher in HBeAg negative group compared with HBeAg positive group,P<0.05;HBV DNA quantity had no statistical importance in each mutant group compared with non-mutant group.Conclusion Several different variant sites of HBV could be detected at one time using the microarray technique.HBV precore region and basic core region mutations is due to HBeAg secretion obstruction in HCC,but has no important effect on the degree of HBV DNA quantity in HCC.