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[摘要] 目的 探讨膀胱移行细胞癌组织中 TIP30/CC3 蛋白的表达及其与患者生物学行为的关系。 方法 选取接受手术治疗的膀胱移行细胞癌(BTCC)存档蜡块 60 例。免疫组化 S-P 法检测 BTCC 标本中 TIP30/CC3 蛋白的表达,并探讨其与患者生物学行为的关系。另取癌旁正常的膀胱组织 10 例(距肿瘤缘 3~5 cm)作对照。 结果 60 例BTCC标本中TIP30/CC3表达阳性19例(31.7%);10 例癌旁正常的膀胱组织 TIP30/CC3表达阳性9 例(90.0%)。BTCC标本中TIP30/CC3阳性率低于癌旁正常的膀胱组织(χ2=9.84,P<0.01)。BTCC 组织中TIP30/CC3表达的肿瘤数量、性别、肿瘤是否复发及转移情况等无统计学意义(P>0.05),与 UICC 分期和病理分级密切相关(P<0.05)。结论 BTCC 组织中 TIP30/CC3 的表达下调或失表达极有可能会促成膀胱癌的发生和发展。BTCC 组织中 TIP30/CC3 的表达与UICC分期和病理分级等生物学行为明显相关,可能参与了膀胱癌发展、转移和浸润的过程,可作为判断其浸润深度、术后复发和预后的评估指标。
[关键词] 膀胱移行细胞癌;免疫组化;TIP30/CC3蛋白;生物学行为
[中图分类号] R737.14 [文献标识码] A [文章编号] 1673-9701(2016)36-0029-03
[Abstract] Objective To investigate the expression of TIP30/CC3 protein in transitional cell carcinoma of bladder and its relationship with the biological behavior in patients. Methods 60 archival paraffin blocks of bladder transitional cell carcinoma(BTCC) treated with surgical treatment were chosen. The expression of TIP30/CC3 protein in BTCC specimens was detected by immunohistochemistry S-P method, and the relationship between TIP30/CC3 protein expression and the biological behavior was explored. Another 10 cases of normal bladder tissue adjacent to the tumor(3-5 cm away from the tumor margin) as the control group. Results TIP30/CC3 expression was positive in 19 cases of 60 BTCC specimens(31.7%). The positive expression of TIP30/CC3 was found in 9 cases(90.0%) of 10 normal bladder tissues. The positive rate of TIP30/CC3 in BTCC specimens was lower than that in normal bladder tissue(χ2=9.84, P<0.01). The expression of TIP30/CC3 in BTCC tissues was not correlated with the tumor quantity, gender, tumor recurrence and metastasis(P>0.05), but was closely correlated with UICC staging and pathological grade (P<0.05). Conclusion Down-regulation or loss of TIP30/CC3 expression in BTCC tissues may contribute to the development and progression of bladder cancer. The expression of TIP30/CC3 in BTCC is significantly correlated with the biological behavior such as UICC staging and pathological grading, and may be involved in the development, metastasis and infiltration of bladder cancer. The expression of TIP30/CC3 in BTCC tissue can be used as an index to judge the depth of invasion, recurrence and prognosis.
[Key words] Bladder transitional cell carcinoma;Immunohistochemistry;TIP30/CC3 protein;Biological behavior
研究發现肿瘤的发生发展过程是一个多基因参与、受多因素影响的、多阶段的发生、发展过程,与肿瘤细胞的过度生长与增殖密切相关[1]。TIP30/CC3 是一种新发现调肿瘤转移抑制相关基因,能促进肿瘤细胞的凋亡,抑制其转移和血管形成等,在大多数正常成人组织中高表达,而在一些肿瘤组织中不表达或表达较弱[2-5]。TIP30/CC3 的促进肿瘤细胞凋亡效应及肿瘤中的作用越来越受临床重视,使其有望成为肿瘤诊断及治疗的新靶点[6-9]。本研究观察了在膀胱移行细胞癌(bladder transitional cell carcinoma,BTCC)组织中TIP30/CC3蛋白的表达及与生物学行为的关系。现报道如下。 1 材料与方法
1.1 一般资料
选取2012年1月~2016年1月我院泌尿科接受手术治疗的 BTCC 存档蜡块 60 例。纳入患者均具有完整临床资料且术后病理证实为BTCC,术前均未行放、化疗。其中男 42 例,女 18 例;年龄 39~87 岁,平均(70.3±6.1)岁。另取癌旁正常的膀胱组织10例(距肿瘤缘 3~5 cm)作为对照。
1.2 免疫组化
采用S-P法。将标本固定于10%福尔马林液中,石蜡包埋后连续4 μm 切片,采用高温修复抗原,室温自然冷却,正常羊血清37℃ 封闭10 min 以减少非特异性背景,倾去血清分别滴加 TIP30/CC3一抗工作液 4℃ 孵育过夜,余操作均参照说明书进行。采用滴加新鲜配置DAB显色,苏木素衬染,梯度酒精脱水,透明、中性树胶封片和镜检。试剂公司购买的阳性切片作阳性对照,以PBS代替一抗作阴性对照。TIP30/CC3蛋白单克隆抗体、多聚赖氨酸和超敏即用型免疫组化S-P试剂盒(通用型)购买自北京中杉生物公司。
1.3 结果判定
TIP30/CC3免疫组化结果以胞膜或胞质出现棕黄色颗粒为阳性细胞。阴性染色除细胞核染成蓝色外,无棕黄色颗粒。按照高倍镜视野下细胞染色强度与阳性细胞比例进行评分,其中染色强度:无色、浅色、棕黄色和褐色分别为 0、1、2 和 3 分;阳性细胞比例:≤10%、11%~50%、51%~75%和>75%分别为0、1、2 和 3 分。染色强度记分×阳性细胞比例记分≥2分为TIP30/CC3( )。
1.4 统计学处理
应用 SPSS18.0软件,计数资料采用χ2检验,P<0.05为差异有统计学意义。
2 结果
2.1 BTCC 和癌旁正常膀胱组织中TIP30/CC3表达差异
60 例BTCC中TIP30/CC3表达阳性19例(31.7%);10 例癌旁正常膀胱组织TIP30/CC3表达阳性9例(90.0%)。BTCC 标本中TIP30/CC3的阳性率低于癌旁正常膀胱組织(χ2=9.84,P<0.01)。
2.2 BTCC 组织中 TIP30/CC3 表达与生物学行为关系
BTCC中TIP30/CC3表达的肿瘤数量、性别、肿瘤是否复发及转移情况等差异无统计学意义(P>0.05),与 UICC分期和病理分级密切相关(P<0.05)。见表1。
3 讨论
TIP30/CC3 基因是一种新发现的与肿瘤转移抑制相关的基因,其蛋白编码产物 TIP30/CC3(Tat interacting protein 30)蛋白是一种分子量为 30 kDa的转录共分子,其本质是一种丝氨酸、苏氨酸激酶,以细胞辅助因子为个体的具有活化并增高人类免疫缺陷病毒-1(HIV-1)转录,可特异性地提高 HIV-1 增殖调节蛋白 Tat 的转录[10]。研究已证实TIP30/CC3基因可抑制肿瘤细胞生长,促进其凋亡,抑制肿瘤血管形成,从而抑制肿瘤的浸润与转移[11-14]。TIP30/CC3基因在肺癌、黑色素瘤、脑瘤、膀胱癌、乳癌、胃肠道癌和肝癌等多种肿瘤中表达下调[15-17]。研究发现乳腺癌组织中 TIP30/CC3 基因的表达水平与淋巴结转移和血管侵袭等病理特征呈负相关。肝癌细胞中上调 TIP30/CC3 基因的表达水平可明显抑制肝癌细胞的增殖和侵袭,下调TIP30/CC3基因的表达水平可加快癌细胞的生长、增殖和侵袭[18]。TIP30/CC3基因在评价肿瘤的预后及基因的靶向治疗中具有重要意义。
近年来 TIP30/CC3 基因在膀胱癌组织中的表达作用越来越引起临床重视。杨涛等[19]研究发现抑癌基因 TIP30/CC3 基因在膀胱癌组织呈低表达或表达缺失状态可能促进膀胱尿路上皮癌的发生、发展及浸润,在膀胱癌患者的治疗以及预防中需重视 TIP30/CC3基因的表达情况。陕光等[20]研究发现抑癌基因 TIP30/CC3 基因在膀胱癌癌组织中呈低表达或表达缺失状态,可能促进 BTCC 的发生和发展,并参与其浸润转移过程。本研究结果发现 BTCC 标本中TIP30/CC3 的表达阳性率明显低于癌旁正常的膀胱组织,表明 BTCC 组织中 TIP30/CC3 的表达下调或失表达极有可能会促成膀胱癌的发生和发展。TIP30/CC3基因具有促凋亡的作用,能够帮助患者抑制肿瘤的生长,抑制膀胱尿路上皮癌组织中的发生和发展,在对患者临床早期的诊断以及治疗的过程中有指导性的作用。癌旁组织中TIP30/CC3的高表达可抑制膀胱尿路上皮癌的发生和发展,对临床的早诊断及治疗起指导效应。同时研究发现 BTCC 组织中TIP30/CC3的表达与UICC分期和病理分级等生物学行为明显相关,表明TIP30/CC3的的表达与 BTCC的恶性程度与浸润深度呈负相关,可能参与了膀胱癌发展、转移和浸润的过程,可作为判断其浸润深度、术后复发和预后的评估指标。有关TIP30/CC3基因在膀胱癌发生、发展、转移和浸润中的作用和相关机制仍需进一步深入研究探讨,以期为膀胱肿瘤的基因靶向治疗提供理论依据。
总之,BTCC 组织中TIP30/CC3的表达下调或失表达极有可能会促成膀胱癌的发生和发展。BTCC 组织中 TIP30/CC3 的表达与 UICC 分期和病理分级等生物学行为明显相关,可能参与了膀胱癌发展、转移和浸润的过程,可作为判断其浸润深度、术后复发和预后的评估指标。
[参考文献]
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[关键词] 膀胱移行细胞癌;免疫组化;TIP30/CC3蛋白;生物学行为
[中图分类号] R737.14 [文献标识码] A [文章编号] 1673-9701(2016)36-0029-03
[Abstract] Objective To investigate the expression of TIP30/CC3 protein in transitional cell carcinoma of bladder and its relationship with the biological behavior in patients. Methods 60 archival paraffin blocks of bladder transitional cell carcinoma(BTCC) treated with surgical treatment were chosen. The expression of TIP30/CC3 protein in BTCC specimens was detected by immunohistochemistry S-P method, and the relationship between TIP30/CC3 protein expression and the biological behavior was explored. Another 10 cases of normal bladder tissue adjacent to the tumor(3-5 cm away from the tumor margin) as the control group. Results TIP30/CC3 expression was positive in 19 cases of 60 BTCC specimens(31.7%). The positive expression of TIP30/CC3 was found in 9 cases(90.0%) of 10 normal bladder tissues. The positive rate of TIP30/CC3 in BTCC specimens was lower than that in normal bladder tissue(χ2=9.84, P<0.01). The expression of TIP30/CC3 in BTCC tissues was not correlated with the tumor quantity, gender, tumor recurrence and metastasis(P>0.05), but was closely correlated with UICC staging and pathological grade (P<0.05). Conclusion Down-regulation or loss of TIP30/CC3 expression in BTCC tissues may contribute to the development and progression of bladder cancer. The expression of TIP30/CC3 in BTCC is significantly correlated with the biological behavior such as UICC staging and pathological grading, and may be involved in the development, metastasis and infiltration of bladder cancer. The expression of TIP30/CC3 in BTCC tissue can be used as an index to judge the depth of invasion, recurrence and prognosis.
[Key words] Bladder transitional cell carcinoma;Immunohistochemistry;TIP30/CC3 protein;Biological behavior
研究發现肿瘤的发生发展过程是一个多基因参与、受多因素影响的、多阶段的发生、发展过程,与肿瘤细胞的过度生长与增殖密切相关[1]。TIP30/CC3 是一种新发现调肿瘤转移抑制相关基因,能促进肿瘤细胞的凋亡,抑制其转移和血管形成等,在大多数正常成人组织中高表达,而在一些肿瘤组织中不表达或表达较弱[2-5]。TIP30/CC3 的促进肿瘤细胞凋亡效应及肿瘤中的作用越来越受临床重视,使其有望成为肿瘤诊断及治疗的新靶点[6-9]。本研究观察了在膀胱移行细胞癌(bladder transitional cell carcinoma,BTCC)组织中TIP30/CC3蛋白的表达及与生物学行为的关系。现报道如下。 1 材料与方法
1.1 一般资料
选取2012年1月~2016年1月我院泌尿科接受手术治疗的 BTCC 存档蜡块 60 例。纳入患者均具有完整临床资料且术后病理证实为BTCC,术前均未行放、化疗。其中男 42 例,女 18 例;年龄 39~87 岁,平均(70.3±6.1)岁。另取癌旁正常的膀胱组织10例(距肿瘤缘 3~5 cm)作为对照。
1.2 免疫组化
采用S-P法。将标本固定于10%福尔马林液中,石蜡包埋后连续4 μm 切片,采用高温修复抗原,室温自然冷却,正常羊血清37℃ 封闭10 min 以减少非特异性背景,倾去血清分别滴加 TIP30/CC3一抗工作液 4℃ 孵育过夜,余操作均参照说明书进行。采用滴加新鲜配置DAB显色,苏木素衬染,梯度酒精脱水,透明、中性树胶封片和镜检。试剂公司购买的阳性切片作阳性对照,以PBS代替一抗作阴性对照。TIP30/CC3蛋白单克隆抗体、多聚赖氨酸和超敏即用型免疫组化S-P试剂盒(通用型)购买自北京中杉生物公司。
1.3 结果判定
TIP30/CC3免疫组化结果以胞膜或胞质出现棕黄色颗粒为阳性细胞。阴性染色除细胞核染成蓝色外,无棕黄色颗粒。按照高倍镜视野下细胞染色强度与阳性细胞比例进行评分,其中染色强度:无色、浅色、棕黄色和褐色分别为 0、1、2 和 3 分;阳性细胞比例:≤10%、11%~50%、51%~75%和>75%分别为0、1、2 和 3 分。染色强度记分×阳性细胞比例记分≥2分为TIP30/CC3( )。
1.4 统计学处理
应用 SPSS18.0软件,计数资料采用χ2检验,P<0.05为差异有统计学意义。
2 结果
2.1 BTCC 和癌旁正常膀胱组织中TIP30/CC3表达差异
60 例BTCC中TIP30/CC3表达阳性19例(31.7%);10 例癌旁正常膀胱组织TIP30/CC3表达阳性9例(90.0%)。BTCC 标本中TIP30/CC3的阳性率低于癌旁正常膀胱組织(χ2=9.84,P<0.01)。
2.2 BTCC 组织中 TIP30/CC3 表达与生物学行为关系
BTCC中TIP30/CC3表达的肿瘤数量、性别、肿瘤是否复发及转移情况等差异无统计学意义(P>0.05),与 UICC分期和病理分级密切相关(P<0.05)。见表1。
3 讨论
TIP30/CC3 基因是一种新发现的与肿瘤转移抑制相关的基因,其蛋白编码产物 TIP30/CC3(Tat interacting protein 30)蛋白是一种分子量为 30 kDa的转录共分子,其本质是一种丝氨酸、苏氨酸激酶,以细胞辅助因子为个体的具有活化并增高人类免疫缺陷病毒-1(HIV-1)转录,可特异性地提高 HIV-1 增殖调节蛋白 Tat 的转录[10]。研究已证实TIP30/CC3基因可抑制肿瘤细胞生长,促进其凋亡,抑制肿瘤血管形成,从而抑制肿瘤的浸润与转移[11-14]。TIP30/CC3基因在肺癌、黑色素瘤、脑瘤、膀胱癌、乳癌、胃肠道癌和肝癌等多种肿瘤中表达下调[15-17]。研究发现乳腺癌组织中 TIP30/CC3 基因的表达水平与淋巴结转移和血管侵袭等病理特征呈负相关。肝癌细胞中上调 TIP30/CC3 基因的表达水平可明显抑制肝癌细胞的增殖和侵袭,下调TIP30/CC3基因的表达水平可加快癌细胞的生长、增殖和侵袭[18]。TIP30/CC3基因在评价肿瘤的预后及基因的靶向治疗中具有重要意义。
近年来 TIP30/CC3 基因在膀胱癌组织中的表达作用越来越引起临床重视。杨涛等[19]研究发现抑癌基因 TIP30/CC3 基因在膀胱癌组织呈低表达或表达缺失状态可能促进膀胱尿路上皮癌的发生、发展及浸润,在膀胱癌患者的治疗以及预防中需重视 TIP30/CC3基因的表达情况。陕光等[20]研究发现抑癌基因 TIP30/CC3 基因在膀胱癌癌组织中呈低表达或表达缺失状态,可能促进 BTCC 的发生和发展,并参与其浸润转移过程。本研究结果发现 BTCC 标本中TIP30/CC3 的表达阳性率明显低于癌旁正常的膀胱组织,表明 BTCC 组织中 TIP30/CC3 的表达下调或失表达极有可能会促成膀胱癌的发生和发展。TIP30/CC3基因具有促凋亡的作用,能够帮助患者抑制肿瘤的生长,抑制膀胱尿路上皮癌组织中的发生和发展,在对患者临床早期的诊断以及治疗的过程中有指导性的作用。癌旁组织中TIP30/CC3的高表达可抑制膀胱尿路上皮癌的发生和发展,对临床的早诊断及治疗起指导效应。同时研究发现 BTCC 组织中TIP30/CC3的表达与UICC分期和病理分级等生物学行为明显相关,表明TIP30/CC3的的表达与 BTCC的恶性程度与浸润深度呈负相关,可能参与了膀胱癌发展、转移和浸润的过程,可作为判断其浸润深度、术后复发和预后的评估指标。有关TIP30/CC3基因在膀胱癌发生、发展、转移和浸润中的作用和相关机制仍需进一步深入研究探讨,以期为膀胱肿瘤的基因靶向治疗提供理论依据。
总之,BTCC 组织中TIP30/CC3的表达下调或失表达极有可能会促成膀胱癌的发生和发展。BTCC 组织中 TIP30/CC3 的表达与 UICC 分期和病理分级等生物学行为明显相关,可能参与了膀胱癌发展、转移和浸润的过程,可作为判断其浸润深度、术后复发和预后的评估指标。
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