【作者】 刘琳；

【导师】 熊永炎； 曾俊；

【作者基本信息】 武汉大学 ， 病理学与病理生理学， 2004， 硕士

【摘要】 目的：检测多药耐药基因（MDR1）、多药耐药相关蛋白（multidrug resistanceassociated protein，MRP）基因和DNA拓扑异构酶Ⅱ α（TopoisomeraseⅡ α，TopoⅡ α）、突变型p53、Bcl-2、P21^ras蛋白在非小细胞肺癌（non—small cell lung cancer，NSCLC）中的表达，观察上述指标与NSCLC各临床病理因素间的关系，探讨NSCLC原发性多药耐药形成的可能机制。 方法：选择武汉大学中南医院病理科1996～2000年未经放、化疗的手术切除NSCLC病例共113例，采用原位分子杂交法检测癌组织中MDR1、MRP基因mRNA的表达，免疫组化方法检测TopoⅡ α、突变型p53、Bcl-2和P21^ras蛋白的表达。 结果：（1） MDR1和MRP mRNA在113例NSCLC组织中的阳性表达率分别为51.3％（58／113）和80.5％（91／113）。不同组织学类型NSCLC组织中，MRP mRNA阳性表达顺序为腺鳞癌＞腺癌＞鳞癌，差异有显著性意义（x²=13.733，P＜0.01）；MDR1mRNA在不同组织学类型中表达无明显差异（P＞0.05）。二者表达与NSCLC的分化程度、淋巴结转移、TNM分期等均无关。MDR1 mRNA和MRP mRNA的协同阳性表达（MDR1⁺／MRP⁺）率为48.7％（55／113）。在不同分化程度鳞癌中，MDR1⁺／MRP⁺表达顺序为中等分化鳞癌＞高分化鳞癌＞低分化鳞癌；不同分化程度腺癌组织中表达顺序为高分化腺癌＞中等分化腺癌＞低分化腺癌；但上述差异均无统计学意义（P＞0.05）。（2） 113例NSCLC组织中TopoⅡ α阳性表达率为60.2％（68／113）。TopoⅡ α在鳞癌中的表达明显高于腺癌和腺鳞癌，差异有显著性意义（x²=6.598，P＜0.05）；而与NSCLC的分化程度、有无淋巴结转移、TNM分期等均无明显相关（P＞0.05）。（3） 113例NSCLC中，突变型p53、Bcl-2和P21^ras阳性表达率分别为61.9％（70／113）、59.3％（67／113）和70.8％（80／113）。p53和P21^ras的表达与NSCLC组织学类型均无明显相关。Bcl-2在不同组织学类型NSCLC中表达顺序为鳞癌＞腺鳞癌＞腺癌，差异具有显著性（x²=15.689，P＜0.01）；经两两比较发现鳞癌中Bcl-2的表达明显高于腺癌（x²=15.295，P＜0.01）。p53的阳性表达率随肿瘤分化程度的升高而递减，差异有显著性（x²=7.690，P＜0.05），但其表达与有无淋巴结转移和TNM分期无关（P＞0.05）。而Bcl-2和P21^ras表达与肿瘤分化程度、有无淋巴结转移和TNM分期等临床病理因素均无明显相关关系（尸>0.05）。（3）NSCLC中MDRI mRNA与MRP mRNA的表达之间存在明显相关（xZ一1 5.535，尸<0.01）。PZz『aS与MnRlm洲A（xZ一20.498，尸<0.01）、P21raS与M砂mRNA（ xZ=8 .486，p一0.004）表达之间有显著相关性。p53和Bcl一2表达之间具有相关性（xZ=4 .697，尸< 0.05）。 结论:（1）非小细胞肺癌原发性多药耐药的形成是一个多因素、多步骤的复杂过程，与包括MDRI、MRP、Topell。及凋亡相关基因等在内的多项因素的参与有关。 （2） MDRI和M即是NSCLC原发性多药耐药形成机制中的重要因素，且MRP所起作用较MDRI更为明显。二者的联合检测更有利于准确判断NSCLC的多药耐药状态。（3） ToPon。的表达减少和/或活性降低与NSCLC的原发性多药耐药形成有一定关系。M对低表达和ToPon。高表达可能是肺鳞癌具有较高化疗敏感性的重要理论依据。（4） NSCLC中MDRI与MRP的表达可能存在某些共同调控机制，PZI『as可能是其共同调控机制之一，而p53和Bcl一2突变介导的耐药机制可能不是通过对MDRI、M妙的调控来实现的。更多还原

【Abstract】 Object: To evaluate expressions of multidrug resistance gene (MDR1), multidrug resistance related protein (MRP) gene, Topoisomerase II a (Topo II a ), mutant p53 protein, Bcl-2 and P21rasin non-small cell lung cancer(NSCLC) of different histological types and different differentiated degrees, lymph node metastasis and TNM staging, and investigate the relationship involved in these characters. Try to elucidate the probable mechanism of intrinsic multidrug-resistance of NSCLC.Methods: NSCLC from 113 untreated cases stored in Department of Pathology, Zhongnan Hospital, Wuhan University between 1996 and 2000 were available for this study. Expressions of MDR1 and MRP gene mRNA in paraffin-embedded cancer tissues were determined by in situ hybridization. And expressions of Topo II a , mutant p53 protein, Bcl-2 and P21ras were detected by S-P immunohistochemical method.Results: (l)The positive expressions of MDR1 and MRP were observed in 58 (51.3%) cases and 91(80.5%) cases of non-small cell lung cancer respectively. The positive expression of MRP in squamous carcinoma was significantly lower than in other histological subtypes ( x2=13.733, P<0.01) , while such difference was not found in the expression of MDR1. Both of MDR1 and MRP had no relationships with cell differentiation, lymphatic metastasis and TNM staging. The positive co-expression of MDR1 and MRP (MDRl+/MRP+)was observed in 55 (48.8%) cases. In squamous carcinoma, the expression of MDR1+/MRP+ in middling differentiated was higher than in high and low differentiated, While the expression of MDR1+/MRP+ in high differentiated adenocarcinoma was the highest in different differentiated adenocarcinoma. And all of them had no statistic differences.(2) The positive percentage of Topo II was 60.2% (68/113) . The expression of Topo II a was significantly related with the histological types but not with differentiated degrees, lymph node metastasis and TNM staging. Topo II a in squamous carcinoma was expressed more highly than in other types of NSCLC (x2=6.598, P<0.05).(3) The positive percentages of mutant p53 protein, Bcl-2 and P21ras were 61.9% (70/113), 59.3% (67/113) and 70.8% (80/113) respectivly. The positive expression of Bcl-2 had a close relationship with the histological types ( x2=15.689, P<0.01) , especially the expression of Bcl-2 in squamous carcinoma was significant higher than in adenocarcinoma ( x2= 15.295, P<0.01) . All the three proteins had no observed relationship with cell differentiation, TNM staging and lymphatic metastasis. (3) There was a significant correlation between expressions of MDRl and MRP ( x2=15.535, P<0.01). The expression of P21ras had close relationships with MDR1( x2 =20.498 , P<0.01) and MRP (zx2=8.486, P=0.004). There was correlation between p53 and Bcl-2 ( x2=4.697, P<0.05).Conclusions: (l)The multridrug of NSCLC is determined by multiple factors and procedures, which include of MDRl, MRP, Topo II a , apoptosis-related gene, et al. (2)Both of MDRl and MRP play significant roles in intrinsic multidrug resistance of non-small cell lung cancer. And MRP gene would be more important than MDRl. The co-evaluate the expressions of MDRl and MRP may help to forecast the instrict multidrug condition of NSCLC better. (3)The reduced expression of Topo II a have relationship with the instrict multidrug of NSCLC. The expressions of MRP and Topo II a are two independent markers which are related with the chemotherapeutic sensitivity in squamous carcinoma. (4) P21ras, instead of P53 and Bcl-2, may be one of the agents that control the expressions of MDRl and MRP genes in multidrug resistance of NSCLC.更多还原