肿瘤抑制基因LASS2在裸鼠膀胱癌模型中的表达及其与肿瘤增殖和凋亡的关系
发布时间：2018-08-13 08:32
【摘要】：目的膀胱癌是泌尿系统最常见的恶性肿瘤之一,关于膀胱癌的发病机制仍未得到全面而深入的研究。LASS2是我国新近发现的肿瘤抑制基因,文中通过检测LASS2在裸鼠膀胱癌模型中的表达,探讨LASS2与肿瘤增殖和凋亡的关系及可能的分子机制。方法采用抽签方法将30只裸鼠随机分为皮下种植组、原代灌注组及空白对照组(DMEM膀胱癌细胞培养基),每组10只。皮下膀胱癌种植模型将已制备的细胞悬液按每只0.5 mL(即5×10~6/0.1 mL个细胞)注射入裸鼠左腋皮下。原位膀胱癌种植模型在裸鼠排空膀胱后将100μL EJ(即1×10~7个细胞)单细胞悬液灌注入膀胱,观察裸鼠致瘤情况。检测不同部位成瘤组织中LASS2、Ki-67的表达及增殖、凋亡等指标变化。结果皮下种植组皮下肿瘤形成,成瘤率为100%。原位灌注组、皮下种植组及空白对照组裸鼠肉眼及病理切片均未检测到肝、肺、肾等器官及淋巴结转移。与空白对照组LASS2表达(81.0%)比较,原位灌注组、皮下种植组LASS2表达(60.0%、14.0%)显著减少(P0.05);与空白对照组Ki-67表达(16.0%)比较,原位灌注组、皮下种植组Ki-67表达(50.0%、78.0%)增加(P0.05);与原位灌注组比较,皮下种植组LASS2表达(14.0%)明显减少(P0.05),Ki-67表达(78.0%)增加(P0.05)。与空白对照组比较,皮下种植组、原位灌注组Bcl-2表达明显升高(P0.05);与皮下种植组比较,原位灌注组Bcl-2表达升高(P0.05);而Bcl-xl在原位种植瘤组织中的表达高于其他两组(P0.05);Bax和Bim均呈现低表达,Bax在各组中的表达水平差异无统计学意义(P0.05);与空白对照组比较,皮下种植组Bim表达明显降低(P0.05),而原位种植瘤组差异无统计学意义(P0.05)。caspase3在各组组织中的表达差异无统计学意义(P0.05);而ki-67在皮下种植瘤组织中的表达较其他两种肿瘤组织中的表达显著升高(P0.05);LASS2在空白对照组中的表达较其他两组表达显著升高(P0.05)。结论 LASS2具有抑制肿瘤增殖的作用,其表达可能与膀胱癌EJ细胞体内成瘤能力、增殖和凋亡有关。
[Abstract]:Objective bladder cancer is one of the most common malignant tumors in the urinary system. The pathogenesis of bladder cancer has not been fully and thoroughly studied. LASS2 is a newly discovered tumor suppressor gene in China. By detecting the expression of LASS2 in nude mice bladder cancer model, the relationship between LASS2 and tumor proliferation and apoptosis and its possible molecular mechanism were studied. Methods by drawing lots, 30 nude mice were randomly divided into subcutaneous implantation group, primary perfusion group and blank control group (DMEM bladder cancer cell culture medium) with 10 in each group. The subcutaneous bladder cancer implantation model injected the prepared cell suspensions into the left axillary subcutaneous of nude mice in 0.5 mL (5 脳 10 ~ (6) / 0. 1 mL cells). In situ bladder cancer implantation model, 100 渭 L EJ (1 脳 107 cells) single cell suspension was perfused into the bladder after bladder emptying in nude mice, and the tumorigenesis of nude mice was observed. The expression, proliferation and apoptosis of LASS2 Ki-67 were detected in different tumor tissues. Results the subcutaneous tumor was formed in the subcutaneous implantation group, and the tumorigenesis rate was 100%. In situ perfusion group, subcutaneous implantation group and blank control group, liver, lung, kidney and lymph node metastasis were not detected in naked eyes and pathological sections of nude mice. Compared with the blank control group (81.0%), the expression of LASS2 in the subcutaneous implantation group (60.0%) decreased significantly (P0.05), and compared with the blank control group (16.0%), the expression of Ki-67 increased (50.0%) in the subcutaneous implantation group (P0.05), and increased in the subcutaneous implantation group (78.0%) (P0.05), compared with the control group (16.0%). The expression of LASS2 (14.0%) in subcutaneous implant group was significantly decreased (P0.05) and Ki-67 expression (78.0%) was increased (P0.05). Compared with the blank control group, the expression of Bcl-2 in the subcutaneous implantation group and in situ perfusion group was significantly increased (P0.05), and compared with the subcutaneous implantation group, The expression of Bcl-2 in situ perfusion group was higher than that in the other two groups (P0.05), and there was no significant difference in the expression of Bcl-xl between the two groups (P0.05). The expression of Bim in subcutaneous implantation group was significantly decreased (P0.05), but there was no significant difference in expression of caspase3 in subcutaneous implanted tumor group (P0.05), while the expression of ki-67 in subcutaneous implanted tumor tissue was higher than that in other two kinds of tumor tissues (P0.05). The expression of LASS2 in the blank control group was significantly higher than that in the other two groups (P0.05). Conclusion LASS2 can inhibit tumor proliferation, and its expression may be related to tumor formation, proliferation and apoptosis of bladder cancer EJ cells.
【作者单位】： 昆明医科大学第二附属医院泌尿外科;
【基金】：国家自然科学基金(81460384) 云南省博士新人奖项目(60116090706) 云南省教育厅科学研究基金(2014J045)
【分类号】：R-332;R737.14
本文编号：2180438
[Abstract]:Objective bladder cancer is one of the most common malignant tumors in the urinary system. The pathogenesis of bladder cancer has not been fully and thoroughly studied. LASS2 is a newly discovered tumor suppressor gene in China. By detecting the expression of LASS2 in nude mice bladder cancer model, the relationship between LASS2 and tumor proliferation and apoptosis and its possible molecular mechanism were studied. Methods by drawing lots, 30 nude mice were randomly divided into subcutaneous implantation group, primary perfusion group and blank control group (DMEM bladder cancer cell culture medium) with 10 in each group. The subcutaneous bladder cancer implantation model injected the prepared cell suspensions into the left axillary subcutaneous of nude mice in 0.5 mL (5 脳 10 ~ (6) / 0. 1 mL cells). In situ bladder cancer implantation model, 100 渭 L EJ (1 脳 107 cells) single cell suspension was perfused into the bladder after bladder emptying in nude mice, and the tumorigenesis of nude mice was observed. The expression, proliferation and apoptosis of LASS2 Ki-67 were detected in different tumor tissues. Results the subcutaneous tumor was formed in the subcutaneous implantation group, and the tumorigenesis rate was 100%. In situ perfusion group, subcutaneous implantation group and blank control group, liver, lung, kidney and lymph node metastasis were not detected in naked eyes and pathological sections of nude mice. Compared with the blank control group (81.0%), the expression of LASS2 in the subcutaneous implantation group (60.0%) decreased significantly (P0.05), and compared with the blank control group (16.0%), the expression of Ki-67 increased (50.0%) in the subcutaneous implantation group (P0.05), and increased in the subcutaneous implantation group (78.0%) (P0.05), compared with the control group (16.0%). The expression of LASS2 (14.0%) in subcutaneous implant group was significantly decreased (P0.05) and Ki-67 expression (78.0%) was increased (P0.05). Compared with the blank control group, the expression of Bcl-2 in the subcutaneous implantation group and in situ perfusion group was significantly increased (P0.05), and compared with the subcutaneous implantation group, The expression of Bcl-2 in situ perfusion group was higher than that in the other two groups (P0.05), and there was no significant difference in the expression of Bcl-xl between the two groups (P0.05). The expression of Bim in subcutaneous implantation group was significantly decreased (P0.05), but there was no significant difference in expression of caspase3 in subcutaneous implanted tumor group (P0.05), while the expression of ki-67 in subcutaneous implanted tumor tissue was higher than that in other two kinds of tumor tissues (P0.05). The expression of LASS2 in the blank control group was significantly higher than that in the other two groups (P0.05). Conclusion LASS2 can inhibit tumor proliferation, and its expression may be related to tumor formation, proliferation and apoptosis of bladder cancer EJ cells.
【作者单位】： 昆明医科大学第二附属医院泌尿外科;
【基金】：国家自然科学基金(81460384) 云南省博士新人奖项目(60116090706) 云南省教育厅科学研究基金(2014J045)
【分类号】：R-332;R737.14
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