低位直肠癌淋巴结转移动物模型的建立及其E-钙黏蛋白的表达

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目的建立裸鼠低位直肠癌淋巴结转移模型,检测E-钙黏蛋白在裸鼠低位直肠癌淋巴结转移模型中的表达,探讨其在直肠癌淋巴结转移过程中的作用。方法取对数生长期的人大肠癌细胞株SW480接种于裸鼠直肠黏膜下,建立裸鼠低位直肠癌淋巴结转移模型。按随机数字表法分为三组:直肠癌模型组(A组,40只,二甲基肼注射+SW480种植)、二甲基肼对照组(B组,40只,二甲基肼注射)和空白对照组(C组,40只,不作任何处理)。处死裸鼠,光镜下观察种植瘤组织和淋巴结转移情况,并采用免疫组化法检测种植瘤组织和转移淋巴结组织中E-钙黏蛋白表达水平。结果 A组裸鼠约1周时成瘤率为100%,之后瘤结节逐渐长大。B组和C组裸鼠均未成瘤。A组裸鼠的肿瘤组织中E-钙黏蛋白阳性表达率(40.0%)明显低于肿瘤周边组织(82.5%),差异有统计学意义(χ2=7.24,P<0.05)。B组裸鼠的直肠黏膜组织中E-钙黏蛋白阳性表达率为80.0%,C组裸鼠的直肠黏膜组织中E-钙黏蛋白阳性表达率为85.0%,A组肿瘤组织以及B组、C组三组比较差异有统计学意义(χ2=5.18,P<0.05),A组的E-钙黏蛋白阳性表达率明显低于B组和C组(χ2=8.12、9.04,P<0.05)。发生侧方淋巴结转移的A组裸鼠种植瘤组织中E-钙黏蛋白阳性表达率为33.3%,明显低于未发生侧方淋巴结转移的裸鼠种植瘤组织(100.0%),差异有统计学意义(χ2=10.28,P<0.05)。结论二甲基肼预处理后进行人大肠癌细胞株SW480接种制模法成功构建了裸鼠低位直肠癌淋巴结转移模型。E-钙黏蛋白表达减弱可能在直肠癌发生、侵袭及淋巴结转移演进过程中起着重要作用。 Objective To establish a lymph node metastasis model of low rectal cancer in nude mice and detect the expression of E-cadherin in lymph node metastasis of low rectal cancer in nude mice and to explore its role in lymph node metastasis of rectal cancer. Methods The logarithmic growth phase human colorectal cancer cell line SW480 was inoculated into the rectal mucosa of nude mice to establish a lymph node metastasis model of low rectal cancer in nude mice. According to random number table, the patients were divided into three groups: rectal cancer model group (group A, 40, dimethyl hydrazine injection + SW480), dimethylhydrazine control group (group B, 40, dimethylhydrazine injection) And blank control group (C group, 40, without any treatment). The nude mice were sacrificed and the implanted tumor tissues and lymph node metastasis were observed under light microscope. The expression of E-cadherin in the implanted tumor and metastatic lymph nodes was detected by immunohistochemistry. Results The nude mice in group A had a tumorigenic rate of 100% at about 1 week, and then the nodules gradually grew up. B group and C group nude mice were not tumor. The positive expression rate of E-cadherin in tumor tissues of group A was significantly lower than that of tumor surrounding tissues (82.0% vs 40.0%, χ2 = 7.24, P <0.05). The positive expression rate of E-cadherin in rectal mucosa of group B was 80.0%, the positive expression rate of E-cadherin in rectal mucosa of group C was 85.0%, the tumor tissue of group A and the group B, The positive expression rate of E-cadherin in group A was significantly lower than that in group B and group C (χ2 = 8.12, 9.04, P <0.05). There was significant difference between group C and group C (χ2 = 5.18, . The positive expression rate of E-cadherin in the nude mice in group A with lateral lymph node metastasis was 33.3%, which was significantly lower than that in the nude mice without lateral lymph node metastasis (100.0%), the difference was statistically significant Significance (χ2 = 10.28, P <0.05). Conclusion The model of lymphatic metastasis of low rectal cancer in nude mice was constructed successfully by inoculation of human colorectal cancer cell line SW480 after dimethylhydrazine pretreatment. Weakening of E-cadherin may play an important role in the development of rectal cancer, invasion and lymph node metastasis.
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