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目的探讨甲基化敏感性高分辨率熔解曲线分析(MS-HRM)在遗传性非息肉性大肠癌(HNPCC)筛查中的应用价值。方法采用实时荧光定量PCR技术检测miR-195在41例散发性大肠癌(SCRC)和9例HNPCC患者癌组织及对应癌旁正常组织(距离癌组织>5 cm)中的表达水平,随后使用MS-HRM检测miR-195启动子区域CpG岛甲基化情况。结果 miR-195在HNPCC癌组织中的表达量为1.20±1.48,甲基化比例为55.56%(5/9);miR-195在SCRC组织中的表达量为0.76±1.06,甲基化比例为58.54%(24/41),差异无统计学意义(P>0.05);miR-195在肠癌组织及癌旁正常组织中的平均表达水平分别为0.837±1.145和2.236±2.468,差异具有统计学意义(P<0.05)。结论 miR-195在SCRC和HNPCC癌组织中的表达有无差异尚待进一步研究。miR-195可能有助于抑制肠癌发展,并且因其甲基化而参于大肠癌的发病机制。
Objective To investigate the value of methylation-sensitive high resolution melting curve analysis (MS-HRM) in the screening of hereditary nonpolyposis colorectal cancer (HNPCC). Methods Real-time fluorescence quantitative PCR was used to detect the expression of miR-195 in 41 cases of sporadic colorectal cancer (SCRC) and 9 cases of HNPCC (corresponding to> 5 cm away from cancer), followed by MS -HRM detection of CpG island methylation in miR-195 promoter region. Results The expression of miR-195 in HNPCC was 1.20 ± 1.48 and the methylation was 55.56% (5/9). The expression of miR-195 in SCRC was 0.76 ± 1.06, and the methylation was 58.54% (24/41) respectively, with no significant difference (P> 0.05). The average expression levels of miR-195 in colorectal cancer tissues and adjacent normal tissues were 0.837 ± 1.145 and 2.236 ± 2.468, respectively Significance (P <0.05). Conclusion Whether miR-195 is differentially expressed in SCRC and HNPCC remains to be further studied. miR-195 may help suppress the development of colorectal cancer, and its methylation may be involved in the pathogenesis of colorectal cancer.