【作者】 韩永华；

【导师】 吴小候；

【作者基本信息】 重庆医科大学 ， 外科学， 2002， 硕士

【摘要】 目的：探讨CK20在膀胱移行细胞癌患者尿脱落细胞和外周血的表达及其意义。方法：采用逆转录聚合酶链反应（RT-PCR）检测：1.膀胱癌细胞株T24、EJ-30 和BIU-87 CK20表达 。2. 61例TCCB患者和11例健康志愿者以及19例非肿瘤患者尿脱落细胞CK20表达。3. 63例TCCB患者和19例健康志愿者以及19例非肿瘤患者外周血CK20表达。以GAPDH作为内参照。结果：1. 膀胱癌细胞株T24、EJ-30 和BIU-87 CK20表达均为阳性。2. 61例 TCCB患者中52例尿脱落细胞CK20阳性表达，阳性率为85.3%，对照组30例中2例CK20阳性表达，阳性率为6.7%，肿瘤组与对照组CK20阳性率有统计学差异（P=0.001）。检测尿脱落细胞CK20表达诊断TCCB，敏感性为85.3%，特异性为93.3%。按临床分期尿脱落细胞CK20表达率T1期为83.9%（26/31），T2-4期 86.7%（26/30），尽管T2-4期表达率稍高于T1期，但两者差异无统计学意义(P>0.05)。按病理分级尿脱落细胞CK20表达率G1为82.8%（24/29），G2 85%（17/20），G3 91.7%（11/12），尽管随分级增加CK20表达率增加，但三者之间CK20表达率差异无统计学意义(P>0.05)。对51例行保留膀胱手术的患者复查尿脱落细胞CK20表达，其中42例术前尿脱落细胞CK20阳性表达者术后阳性表达6例，阳性表达率为14.3%（6/42），9例术前尿脱落细胞CK20阴性表达者术后均为阴性表达。本组有29例患者获得随访6个月，其中术后尿脱落<WP=6>细胞CK20阳性表达4例，阴性表达25例，4例CK20阳性表达者有2例（50%）复发，25例CK20阴性表达者有1例（4%）复发，两者复发率有统计学差异（P=0.04）。3. 19例健康志愿者和19例非肿瘤患者外周血CK20的表达均为阴性；63例TCCB患者外周血CK20阳性表达率为25.4%（16/63），其中T1期为0（0/31），T2期为12.5%（2/16），T3期为77.8%（7/9），T4期为100%（7/7），CK20阳性表达率随分期增加而增加，T3-4期明显高于T1期和T2 期（P<0.05）。对T1-3期33例患者随访6个月，其中外周血CK20阴性表达26例（T1 期18例， T2 期8例），随访期间未发现远处转移；外周血CK20阳性表达7例（T2 期2例，T3 期5例）中2例发生远处转移，其中骨转移1例，腹膜后淋巴结转移1例。结论：1.检测尿脱落细胞CK20表达诊断TCCB敏感性和特异性高，可作为TCCB早期诊断以及术后监测的方法之一。2.检测外周血CK20表达，可以提示癌细胞的血行播散，对判断预后以及指导治疗具有一定临床价值。更多还原

【Abstract】 Objective To determine if detection of CK20 expression in urine and peripheral blood by using RT-PCR can provide a novel noninvasive tool for the diagnosis of transitional cell carcinoma of the bladder .Methods CK20 mRNA in three bladder cancer cell lines (T24 , EJ-30 and BIU-87) , exfoliated urinary cells of 61 patients with TCCB, 19 non-cancerous patients and 11 healthy volunteers and peripheral blood of 63 patients with TCCB, 19 non-cancerous patients and 19 healthy volunteers was detected. GAPDH was used as a general marker for normal urinary cells and normal blood cells. CK20 and GAPDH were amplified by using RT-PCR. Results 1.The positive rate of CK20 in bladder cancer cell lines was 100% (3/3). 2. Only 2 of 30 (6.7%) showed positive CK20 signals in urine from control group. Fifty-two of 61 (85.3%) patients with TCCB showed positive CK20 signals in urine. The CK20 expression difference between experimental group and control group was significant (p<0.05). The positive rate of CK20 in T1 was 83.9%（26/31），T2-4 86.7%（26/30）；G1 82.8%（24/29），G2 85%（17/20），G3 91.7%（11/12）。Six of 42 preoperative CK20-positive patients showed positive<WP=8>CK20 signals in urine after tumor removal. Nine of 9 preoperative CK20-negative patients showed negative CK20 signals in urine after tumor removal. Two of 4 postoperative CK20-positive patients suffered from recurrence, only 1 of 25 postoperative CK20-negative patients suffered from recurrence during a 6-month follow-up period (p=0.04). 3. Nineteen healthy volunteers and 19 non-cancerous patients showed no CK20 signals in peripheral blood. Sixteen of 63 (25.4%) patients with TCCB showed positive CK20 signals in peripheral blood, including 0/31 of stage T1, 2/16 of stage T2, 7/9 of stage T3 , 7/7 of stage T4; The CK 20 expression difference between T1 / T2 group and T3-4 groups was significant (p<0.05). Two patients with positive CK20 signals in peripheral blood had distant metastases during a 6-month follow-up period.Conclusions CK20 is applicable to detect cancer cells in urine and peripheral blood of patients with TCCB and may be helpful in early diagnosis and follow-up of TCCB.更多还原