【作者】 李俊；

【导师】 张旭；

【作者基本信息】 华中科技大学 ， 外科学， 2010， 博士

【摘要】 第一部分p73、MGMT基因在膀胱尿路上皮癌中的表达及其临床意义目的研究p73、MGMT基因在膀胱尿路上皮癌标本组织中的表达情况及其与临床病理特征的关系。方法应用免疫组织化学SP法和逆转录—聚合酶链反应(RT-PCR)技术检测p73、MGMT基因在60例膀胱尿路上皮癌组织标本中的表达,统计分析其与肿瘤临床病理的关系。结果免疫组化显示,膀胱癌组织中p73的阳性表达率为38.3%(23/60),明显低于癌旁组织(78.3%,47/60)及正常膀胱组织(86.7%,13/15),p<0.01；P73蛋白的表达与膀胱尿路上皮癌组织分化程度有关(p<0.05),且与临床分期、浸润及转移也密切相关(p<0.05),MGMT在膀胱癌组织中的蛋白表达率为35.0%,明显低于癌旁组织(76.7%)及正常膀胱组织(86.7%),在高级别(G3)膀胱组织中的MGMT阳性表达率为11.8%,低于低级别(G1～2)膀胱癌组织44.2%的阳性表达率,但发现MGMT蛋白的阳性表达率在膀胱癌的临床分期的发生发展中却无明显统计学差异；RT-PCR显示,p73 mRNA在肿瘤组织中的阳性表达率为53.4%,与正常膀胱组织和癌旁组织标本相比表达明显下降,在不同临床分级肿瘤组织中,高级别(G3)膀胱癌组织中p73基因mRNA的阳性表达率为17.6%,远低于低级别(G1～2)膀胱癌组织,而且在不同临床分期的肿瘤组织中,肌层浸润性膀胱癌组织中p73基因mRNA的阳性表达率(22.7%)与非肌层浸润性膀胱癌组织中的也有明显统计学差异,MGMT mRNA在肿瘤组织中的阳性表达率为38.3%,与正常膀胱组织和癌旁组织标本相比表达明显下降,在不同临床分级肿瘤组织中,高级别(G3)膀胱癌组织中MGMT基因mRNA的阳性表达率为17.6%,也要低于低级别(G1～2)膀胱癌组织。而在不同临床分期的肿瘤组织中,肌层浸润性膀胱癌组织中MGMT基因mRNA的阳性表达率(31.8%)与非肌层浸润性膀胱癌组织中的无明显统计学差异。结论p73、MGMT基因可能在尿路上皮癌的发生和发展中起起到一定抑制作用；p73蛋白和MGMT蛋白的的表达有可能成为一种判断尿路上皮癌预后的重要的肿瘤标记物之一。第二部分p73、MGMT基因启动子甲基化与膀胱尿路上皮癌生物学行为关系的研究目的检测膀胱尿路上皮癌组织和外周血中p73基因和O6-甲基鸟嘌呤-DNA甲基转移酶(MGMT)启动子甲基化状态,探讨p73、MGMT甲基化与其蛋白表达水平以及肿瘤生物学行为之间的关系。方法应用甲基化特异性PCR(MSP)法分别检测60例膀胱尿路上皮癌患者的肿瘤组织和外周血中及15例正常膀胱粘膜组织中p73、MGMT基因启动子的甲基化状态。结果膀胱尿路上皮癌组织中p73甲基化阳性率为43.3%(26/60),外周血中p73甲基化阳性率为40.0%(24/60),明显高于正常膀胱组织(0.0%,0/15),p<0.01；p73启动子甲基化与其表达呈负相关,p<0.05；高级别(G3)膀胱癌组织中p73甲基化阳性率要比低级别(G1-2)膀胱癌高,p<0.05,并且肌层浸润性(T2-4)膀胱癌组织中p73甲基化阳性率也要比非肌层浸润性(Ta～1)膀胱癌组织中高；同期实验还表明膀胱癌组织中MGMT甲基化阳性率为45.0%(27/60),外周血中MGMT甲基化阳性率为40.0%(24/60),明显高于正常膀胱组织(0.0%,0/15),p<0.01;MGMT启动子甲基化与其表达呈负相关,p<0.05；高级别(G3)膀胱癌组织和外周血中MGMT甲基化阳性率要比低级别(G1～2)膀胱癌高,p<0.05,而MGMT甲基化与膀胱癌临床分期却无明显关系。结论p73和MGMT基因启动子甲基化可能参与了膀胱尿路上皮癌的发生、发展和肿瘤分化；MGMT启动子的高甲基化有望成为膀胱尿路上皮癌早期诊断和预后判断的一种重要肿瘤标记物之一。目的探讨后腹腔镜手术治疗肾上腺囊肿的方法及临床意义。方法2000年3月～2008年7月我们采用后腹腔镜手术治疗了27例肾上腺囊肿。扩张后腹膜间隙后,紧靠膈下纵形切开肾周筋膜,分离肾上腺周围三个相对无血管间隙,游离肾上腺后21例行肾上腺囊肿部分切除术,6例行肾上腺全切除术。结果所有手术均获成功,手术时间(35.1±11.6)min,术中出血量(27.9±13.6)m1,住院时间(4.3±1.8)d。术后无严重并发症,随访12～36个月,无一例复发。结论后腹腔镜手术可作为肾上腺囊肿治疗的标准术式。更多还原

【Abstract】 Part One Expression of p73 gene and MGMT gene in urinary epithelia carcinoma of bladder and its clinical significanceObjective To investigate the expresion of p73 gene and MGMT gene in urinary epithelia carcinoma of bladder and the relationship between the expression and clinical pathological characteristics.Methods Immunohistochemistry and RT-PCR were used to detect the expression of p73 protein and mRNA and MGMT protein and mRNA in 60 urinary epithelia carcinoma tumor tissue. Correlations of the expression of p73 and MGMT gene to clinicopathologic parameters were statistically analyzed.Results Under the immunohistochemistry study, the positive rate of P73 protein expression in urinary epithelia carcinoma was 38.3%(23/60), which was obviously lower than that in paracarcinoma tisue(78.3%,47/60) and normal bladder tisue(86.7%,13/15) (p<0.01).Furthermore, protein expresion of P73 was significantly correcated with pathological grade of cancers (p<0.05). And statistical significance was also found between the expresion of p73 and clinical stage of the cancers (p<0.05). The positive rate of MGMT protein expression in urinary epithelia carcinoma was 35.0%, which was obviously lower than that in paracarcinoma tissue (76.7%) and normal bladder tissue (86.7%). Furthermore, the positive rate of MGMT protein expression in G3,which was 11.8%, was also lower than that in G1～2 (44.2%).But there was no statistical significance found between the expression of MGMT and clinical stage of urinary epithelia carcinoma; Under the RT-PCR study, the positive rate of p73 mRNA expression in urinary epithelia carcinoma was 53.4%, which was obviously lower than that in paracarcinoma tissue and normal bladder tissue. And the expression of p73 mRNA was significantly decreased in G3 stage tumor, which was 17.6%, to G1～2 stage tumor. And there was also found statistical significance between the expression of p73 mRNA and clinical stage of urinary epithelia carcinoma. The positive rate of MGMT mRNA expression in urinary epithelia carcinoma was 38.3%,which was obviously lower than that in paracarcinoma tissue and normal bladder tissue. And the expression of MGMT was significantly decrease in G3 stage tumor, which was 17.6%, to lower ones. But there was also no statistical significance found between the expression of MGMT mRNA and clinical stage of urinary epithelia carcinoma.Conclusions p73 gene and MGMT gene may play important role in carcinogenesis and differentiation of transitional carcinoma of the bladder. Therefore the expresion of p73 and MGMT could be one of prognostic indicator of urinary epithelia carcinoma of bladder. Part Two Relationship between Aberrant Methylation of p73 and MGMT gene Promoter and Biological Behaviour of urinary epithelia carcinoma of bladderObjective To detect the promoter methylation of p73 gene and O6-methylguanine-DNA methyltransferase (MGMT) in urinary epithelia carcinoma of bladder and vein blood samples, analyze the relationship between p73 and MGMT promoter methylation and their protein expression, and also biological behaviour of bladder cancer.Methods Promoter methylation of p73 and MGMT gene was detected by methylation-specific PCR (MSP) in 60 samples of urinary epithelia carcinoma of bladder and vein blood and 15 normal bladder tissue samples. Correlations of the aberrant methylation of p73 and MGMT Promoter to clinicopathologic parameters were statistically analyzed.Results The positive rate of p73 promoter methylation were 43.3% (26/60) in urinary epithelia carcinoma of bladder and 40.0% (24/60) in vein blood, which was obviously higher than that in normal bladder tissue (0.0%,0/15), p<0.01.A significantly relationship was found between the aberrant methylation of p73 and its protein expression, p<0.05;The positive rates of p73 promoter methylation in high-grade and low-grade urothelia carcinoma was different, p<0.05.And statistical significance was also found between the positive rates of p73 and clinical stage of the cancers (p<0.05). In the same time, the positive rate of MGMT promoter methylation were 45.0%(27/60) in urinary epithelia carcinoma and 40.0%(24/60) in vein blood, which was obviously higher than that in normal bladder tissue (0.0%,0/15), p<0.01.A significantly relationship was found between the aberrant methylation of MGMT and its protein expression, p<0.05.Furthermore, the positive rates of MGMT promoter methylation in high-grade and low-grade urothelia carcinoma was different, p<0.05.But no statistical significance was found between the MGMT promoter methylation and clinical stage of bladder cancer.Conclusions p73 gene and MGMT gene promoter hypermethylation play important role in carcinogenesis and differentiation of bladder urothelia carcinoma, which could be one of prognostic indicator of urinary epithelia carcinoma of bladder. Objective To evaluate the operative methods and clinical significance of retroperito-neoscopic operation for adrenal cysts.Methods Clinical data of 27 patients with adrenal cyst treated by retroperitoneoscopic operations in our hospital From March 2000 to July 2008 were analyzed retrospectively. After the retroperitoneal space was dilated, the perinephric fascia was opened longitudinally near the diaphragm. The three non-vascular space relatively around the adrenal gland were dissected, Then 21 cases performed partial adrenalectomy and 7 cases total adrenalectomy.Results All the operations were successfully conducted. The mean operative time was 35.1±11.6 min, the mean blood loss was 27.9±13.6 ml, and the mean hosp ital stay was 4.3±1.8 d. No postoperative comp lications were seen after operation, and follow-up for 12～36 months in all the patients found no recurrence.Conclusions Retroperitoneoscopic operation can be used as the standard treatment of adrenal cyst surgical.更多还原