MTA1靶向shRNA对裸鼠喉鳞癌移植瘤生长和转移能力的影响

发布时间：2018-03-10 21:31

本文选题：喉鳞状细胞癌　切入点：肿瘤转移相关基因1　出处：《重庆医科大学》2015年硕士论文　论文类型：学位论文

【摘要】：目的：喉癌的发病率约是全身恶性肿瘤的5%,是最常见的头颈部癌症之一,其组织学类型95%是鳞状细胞癌。对喉癌患者最终结果的主要影响是肿瘤复发和转移,而淋巴结转移是喉癌患者复发和死亡的常见原因,也是喉癌最重要的转移途径。肿瘤转移相关基因1(metastasis-associated gene 1, MTA1)最初是大鼠乳腺癌转移灶筛选出的肿瘤转移相关基因。其对乳腺癌转移能力有促进作用,因此被命名为肿瘤转移相关基因。在人类恶性肿瘤如肺癌、卵巢癌、胃癌、骨肉瘤等中表达量增加,在mRNA水平的高表达与肿瘤的侵袭和淋巴结转移相关。本试验通过RNA干扰(RNAi)技术,观察肿瘤转移相关基因1对裸鼠喉鳞癌移植瘤生长和转移能力的影响。方法：构建MTA1 shRNA1侵病毒干扰载体,进行包装与转染Hep-2细胞。将阴性对照组和MAT1 shRNA感染的Hep-2细胞接种于裸鼠爪垫,每组5只。复制人喉癌移植瘤淋巴转移模型,9周后处死裸鼠,取下肿瘤组织及腹股沟淋巴结,进行形态学学检测。反转录PCR检测MTA1、β连环素(β-catenin)、基质金属蛋白酶9(MMP-9)、细胞周期蛋白D1(CyclinD1) mRNA的水平,Western blot法检测MTA1、β-catenin、 MMP-9、 CyclinDl蛋白水平。结果：慢病毒MTA1干扰载体筛选成功；2组人喉癌裸鼠爪垫移植瘤100%成瘤：MAT1 shRNA转染细胞裸鼠肿瘤体积在同一时间点均明显小于阴性对照组；MAT1 shRNA转染细胞5只裸鼠爪垫肿瘤未发现淋巴结转移；阴性对照组5只裸鼠腹股沟淋巴结切片均可见喉癌细胞；与阴性对照组相比,MAT1 shRNA转染细胞MTA1、β-catenin、MMP-9、CyclinD1的mRNA和蛋白水平明显降低。结论：抑制MTA1基因能够减缓裸鼠喉鳞癌的生长并降低其转移能力,其机制可能与MTA1影响β-catenin、Cyclin D1、MMP-9的表达水平有关。
[Abstract]:Objective: the incidence of laryngeal cancer is about 5% of systemic malignant tumors, and is one of the most common head and neck cancers. Its histological type 95% is squamous cell carcinoma. The main effect on the final outcome of laryngeal cancer is tumor recurrence and metastasis. Lymph node metastasis is a common cause of recurrence and death in patients with laryngeal cancer. Metastasis-associated gene 1 (Metastasis-associated gene 1, MTA1) is the most important metastasis pathway for laryngeal carcinoma. So it's named tumor Metastasis-Related Gene. Increased expression in Human malignant tumors such as Lung Cancer, Ovarian Cancer, gastric Cancer, Osteosarcoma, etc. High expression of mRNA was associated with tumor invasion and lymph node metastasis. To observe the effect of tumor metastasis related gene 1 on the growth and metastasis ability of transplanted laryngeal squamous cell carcinoma (LSCC) in nude mice. Methods: MTA1 shRNA1 invading virus interference vector was constructed. Hep-2 cells were packaged and transfected. The negative control group and MAT1 shRNA infected Hep-2 cells were inoculated into the paw pad of nude mice with 5 in each group. After 9 weeks of lymphatic metastasis model of human laryngeal carcinoma transplanted tumor, nude mice were killed and tumor tissues and inguinal lymph nodes were removed. The levels of MTA1, 尾 -cateninine (尾 -cateninine), matrix metalloproteinase-9 (MMP-9) and cyclin D _ (1) mRNA were detected by reverse transcription PCR and the levels of MTA1, 尾 -catenin, MMP-9 and CyclinDl were detected by Western blot. Results: lentivirus MTA1 interference vector was screened successfully in two groups. At the same time, the tumor volume in nude mice transfected with 100%% MAT1 shRNA was significantly smaller than that in the negative control group (5 nude mice). No lymph node metastasis was found in the tumor. Laryngeal carcinoma cells were found in the inguinal lymph node sections of 5 nude mice in negative control group. Compared with the negative control group, the levels of mRNA and protein of MTA1, 尾 -cateninin MMP-9 and cyclin D1 were significantly decreased. Conclusion: inhibition of MTA1 gene can inhibit the growth and metastasis of laryngeal squamous cell carcinoma in nude mice, and its mechanism may be related to the effect of MTA1 on the expression of MMP-9 in 尾 -cateninine cyclin D1mRNA.
【学位授予单位】：重庆医科大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R739.65

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