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目的检测乙型肝炎病毒(HBV)相关肝细胞癌(HCC)组织及癌旁组织中β链接素(βcatenin)基因第3外显子突变及其表达异常,探讨其在HBV相关肝细胞癌中的作用。方法采用聚合酶链单链构象多态性(PCRSSCP)、直接测序、逆转录聚合酶链反应(RTPCR)、免疫组织化学技术等方法,检测27例HBV相关肝细胞癌及27例癌旁组织βcatenin基因第3外显子点突变以及βcateninmRNA表达差异性。结果(1)HCC组织中,βcatenin基因第3外显子的点突变率约26%(7/27),分别为密码子37(TCT→TGT),丝氨酸→半胱氨酸;密码子45(TCT→TTT),丝氨酸→苯丙氨酸;密码子35(ATC→ATG),异亮氨酸→蛋氨酸。癌旁组织均未发现突变(P<0.05)。(2)βcateninmRNA在癌组织中的表达量明显高于癌旁组织(0.86±0.07比0.33±0.04,P<0.05)。(3)βcatenin蛋白在癌细胞胞浆及核内聚集。结论βcatenin基因第3外显子突变及βcatenin的过度表达对HBV相关肝细胞癌发生起到重要作用。
Objective To detect the mutation and expression of exon 3 of β-catenin in hepatocellular carcinoma (HCC) tissues and adjacent tissues of hepatitis B virus (HBV) effect. Methods PCR-SSCP, direct sequencing, reverse transcription polymerase chain reaction (RTPCR) and immunohistochemistry were used to detect the expression of βcatenin in 27 HBV-related hepatocellular carcinomas and 27 para-cancerous tissues Point mutation of exon 3 gene and the difference of βcatenin mRNA expression. Results (1) The point mutation rate of βcatenin gene exon 3 in HCC tissues was 26% (7/27), which were codon 37 (TCT → TGT), serine → cysteine, codon 45 ( TCT → TTT), serine → phenylalanine; codon 35 (ATC → ATG), isoleucine → methionine. No changes were found in adjacent tissues (P <0.05). (2) The expression of βcatenin mRNA in cancer tissues was significantly higher than that in adjacent tissues (0.86 ± 0.07 vs. 0.33 ± 0.04, P <0.05). (3) βcatenin protein accumulation in the cytoplasm and nucleus of cancer cells. Conclusion The mutation of exon 3 of β-catenin gene and the overexpression of β-catenin play an important role in the development of HBV-related hepatocellular carcinoma.