胆管细胞癌细胞CD147表达及其临床意义

Expression of CD147 and its clinical significance in patients with intrahepatic cholangiocarcinoma complicating intrahepatic bile duct stones

目的探讨CD147在肝胆管结石并发胆管细胞癌(ICC)组织的表达及其临床意义。方法收集肝胆管结石并发ICC标本30例、对应的癌旁肝组织标本30例和正常肝标本10例。采用q RT-PCR法检测组织CD147m RNA水平,采用免疫组化法检测CD147蛋白的表达;采用Western-Blotting法检测基质金属蛋白酶-2(MMP-2)和9(MMP-9)蛋白的表达;采用肯德尔(Kendall)等级相关分析CD147 m RNA、CD147、MMP-2和MMP-9表达的相关性。结果经q RT-PCR检测显示,肝胆管结石并发ICC组织CD147m RNA水平为(3.67±1.88),显著高于癌旁组织【(1.52±0.57),P<0.01】和正常肝组织【(1.05±0.32),P<0.01】,癌旁组织高于正常肝组织(P<0.01);>5 cm ICC组织CD147 m RNA相对水平(2.73±0.97)显著高于<5 cm肿瘤组织【(1.03±0.32),P<0.01】,有淋巴转移(2.68±0.74)组织显著高于无转移组织【(1.07±0.44),P<0.01】,肿瘤分化程度低组织(2.71±0.86)显著高于中高分化组织【(1.06±0.42),P<0.01】;不同年龄和性别患者ICC组织CD147 m RNA水平无显著性差异(P>0.05);经免疫组化检测显示,肝胆管结石并发ICC组织CD147蛋白表达相对水平为(27.95±4.90),显著高于癌旁组织【(13.34±9.59),P<0.01】和正常肝组织【(2.18±0.66),P<0.01】,而癌旁组织高于正常肝组织(P<0.01);ICC组织MMP2表达量为(1.06±0.13),显著高于癌旁组织【(0.20±0.03),P<0.01】和正常肝组织【(0.15±0.02),P<0.01】,而癌旁组织MMP2表达量显著高于正常肝组织(P<0.01);肝胆管结石并发ICC组织MMP9表达量为(0.93±0.12),显著高于癌旁组织【(0.17±0.03),P<0.01】和正常肝组织【(0.15±0.03),P<0.01】,而癌旁组织MMP9表达量显著高于正常肝组织(P<0.01);在ICC组织中,CD147m RNA水平与MMP2蛋白表达呈正相关(r=0.457,P<0.05);CD147m RNA水平与MMP9蛋白表达也呈正相关(r=0.428,P<0.05);CD147蛋白与MMP2蛋白表达呈正相关(r=0.543,P<0.05),CD147蛋白与MMP9蛋白表达也呈正相关(r=0.517,P=0.05)。结论 CD147 m RNA、CD147、MMP2、MMP9均在肝胆管结石并发ICC组织中表达增强,CD147的表达可能促进了MMP2和MMP9的表达上调,它们可能共同参与了胆管癌的发生和发展过程。

Objective To investigate the expression of CD147 in intrahepatic cholangiocarcinoma（ICC） and its clinical significance. Methods 30 ICC samples with hepatolithiasis,corresponding paracancerous tissues and 10 normal liver tissues were collected in our hospital. The CD147 mRNA in liver tissues was detected by qRT-PCR, and the expression of CD147 protein was detected by immunohistochemical staining. The expression of matrix metalloproteinase -2（MMP-2） and 9 （MMP-9） protein were detected by Western blotting. The correlation of CD147 mRNA,CD147,MMP-2 and MMP-9 expression was analyzed by Kendall. Results qRT-PCR detection showed that CD147 mRNA level in ICC tissues was（3.67±1.88）,much higher than in their corresponding paracancerous tissues【（1.52±0.57）,P〈0.01】 or normal liver tissues【（1.05±0.32）,P〈0.01】,and there was a significant difference between paracancerous tissues and normal liver tissues（P〈0.01）;the relative CD147 mRNA level in 〉5 cm ICC（2.73±0.97） was much higher than in 〈5 cm ICC tissues【（1.03±0.32）,P〈0.01】,with lymph node metastasis（2.68±0.74） was much higher than ICC without【（1.07±0.44）,P〈0.01】,and in ICC with lower differentiation（2.71±0.86） was much higher than in with high differentiation 【（1.06±0.42）,P〈0.01】;there was no significant difference between young and elderly or male and female patients（P〉0.05）; immunohistochemical staining detection showed the CD147 protein expression in ICC tissues was（27.95±4.90）,much higher than in paracancerous tissues【（13.34±9.59）,P〈0.01】 or normal liver tissues【（2.18±0.66）,P〈0.01】,and there was a significant difference between paracancerous tissues and normal liver tissues （P〈0.01）;the MMP2 expression in ICC tissues was （1.06±0.13）,much higher than in paracancerous tissues【（0.20±0.03）,P〈0.01】 or normal liver tissues【（0.15±0.02）,P〈0.01】,and there was also a significant difference between paracancerous tissues and normal liver tissues（P〈0.01）;the MMP9 expression in ICC was （0.93±0.12）,much higher than in paracancerous tissues 【（0.17±0.03）,P〈0.01】 or normal liver tissues 【（0.15±0.03）,P〈0.01】,and there was a significant difference between paracancerous tissues and normal liver tissues（P〈0.01）;the CD147mRNA levels in ICC was positively correlated to MMP2 expression（r=0.457,P〈0.05）,CD147mRNA positively to MMP9 expression （r=0.428,P〈0.05）, CD147 protein expression positively to MMP2 expression（r=0.543,P〈0.05）,and CD147 positively to MMP9 expression（r=0.517,P=0.05）. Conclusions There is a phenomenon of increased CD147 mRNA levels and enhanced expression of MMP2 and MMP9 in ICC tissues,which might promote the carcinogenesis,tumor growth,lymph node metastasis,and the lower tumor differentiation.