原发性肝癌不同血瘀证患者肝组织微小RNA表达差异的初步研究

MicroRNA expression profiles of hepatic tissue in primary hepatic carcinoma patients with different blood stasis syndrome

目的对原发性肝癌中医辨证分型提供客观的量化标准。方法采用微小RNA(miRNA)芯片检测、实时定量PCR验证,对比观察原发性肝癌气虚血瘀证、肝郁血瘀证、肝胆湿热证及肝瘀痰结证患者(各3例,分别以证型命名组名)肝组织10种miRNA表达水平的差异;并与3例正常肝组织(正常组)进行比较。结果 miR-122-3p各证型组均明显高于正常组(P<0.05),气虚血瘀组明显高于其他各证型组(P<0.05);miR-30b-5p在肝胆湿热组明显低于其他各证型组(P<0.05),其他各证型组与正常组比较无显著性差异;miR-182-5p在气虚血瘀组、肝郁血瘀组和肝胆湿热组均明显高于正常组(P<0.05);miR-221-5p在肝胆湿热组和肝郁血瘀组明显低于其他各证型组及正常组(P<0.05),但这两组间无显著性差异;miR-221-3p在肝郁痰结组高于其他各证型组,但无统计学差异;miR-21-5p在气虚血瘀组、肝郁血瘀组和肝郁痰结组均高于肝胆湿热组和正常组,但无统计学差异。miR-222-3p、miR-214-3p、miR-491-3p、miR-422a各组间未见明显差异。结论原发性肝癌不同血瘀证患者肝组织miRNA水平存在显著性差异;本研究为原发性肝癌的中医辨证分型提供了客观的量化标准。

Objective To provide an objective quantitative standard for traditional Chinese medicine syndrome-types of primary hepatic carcinoma. Methods The differences about 10 kinds of miRNA expression in hepatic tissue between qi deficiency and blood stasis syndeome,liver stagnation and blood stasis syndeome,gan-dan damp-heat syndeome,liver stasis phlegm stagnation syndeome patients with hepatocellular carcinoma were compared by using miRNA microarray and RT PCR method. There were 3 cases in each group which named as their syndrome. All the hepatic tissues were compared with3 cases of normal liver tissues（ normal group）. Results MiR-122-3p in hepatic tissue was higher than the normal group（P〈0. 05）. MiR-122-3p in qi deficiency and blood stasis syndeome group was higher than the other group（P〈0. 05）.MiR-30b-5p in gan-dan damp-heat syndeome group was lower than the other three groups（P〈0. 05）,which was similar with the normal group. MiR-182-5p in qi deficiency and blood stasis syndeome group,liver stagnation and blood stasis syndeome group,gan-dan damp-heat syndeome group were higher than normal group（P〈0. 05）. MiR-221-5p in liver stagnation blood stasis syndeome group and gan-dan damp-heat syndeome group were lower than the other groups（P〈0. 05）.But there was no significant difference between the two groups. MiR-221-3p in liver stasis phlegm stagnation syndeome group was higher than the other groups,with no statistical difference. MiR-21-5p in qi deficiency and blood stasis syndeome group,liver stagnation and blood stasis syndeome group were higher than gan-dan damp-heat syndeome group and liver stasis phlegm stagnation syndeome group,but there was no significant difference. There was no significant difference in MiR-222-3p,miR-214-3p,miR-491-3p,miR-422 a of each group. Conclusions MiRNA expression in different kinds of blood stasis syndeome in primary hepatic carcinoma patients was different. This research provide an objective quantitative standard for traditional Chinese medicine syndrome differentiation of primary liver cancer classification.