本文選題：人結(jié)直腸癌 + 淋巴道轉(zhuǎn)移　； 參考：《重慶醫(yī)科大學(xué)》2008年碩士論文

【摘要】： 結(jié)直腸癌是消化道常見腫瘤,惡性程度較高,易發(fā)生淋巴性轉(zhuǎn)移和消化道種植播散。腫瘤浸潤和轉(zhuǎn)移不僅影響著臨床治療效果,也是患者腫瘤復(fù)發(fā)、死亡的主要原因,因而對結(jié)直腸癌轉(zhuǎn)移機(jī)制及其防治對策的研究是當(dāng)前的重要任務(wù)。而人結(jié)直腸癌裸鼠移植轉(zhuǎn)移模型的建立是當(dāng)前國內(nèi)外關(guān)注的課題,為了能更好地研究結(jié)直腸癌轉(zhuǎn)移機(jī)制和防治對策,建立穩(wěn)定、可靠、重復(fù)性好的適合動(dòng)物模型是十分必要的。這可為進(jìn)一步研究結(jié)直腸癌的浸潤轉(zhuǎn)移機(jī)制、藥物干預(yù)等抗轉(zhuǎn)移治療提供有效的實(shí)驗(yàn)平臺,利于改觀結(jié)直腸癌的預(yù)后和結(jié)局。但目前尚未構(gòu)建成功較為理想的結(jié)直腸癌單純性淋巴道轉(zhuǎn)移動(dòng)物模型及結(jié)直腸癌粘膜原位移植轉(zhuǎn)移模型。因此,我們從模型的角度利用人結(jié)腸癌細(xì)胞株HCT-116,研究構(gòu)建人結(jié)直腸癌裸小鼠爪墊單純性淋巴道移植轉(zhuǎn)移動(dòng)物模型,以及組織塊法直腸粘膜原位移植轉(zhuǎn)移動(dòng)物模型的可行性具有重要的科研價(jià)值和臨床意義。 第一部分人結(jié)直腸癌裸小鼠淋巴道轉(zhuǎn)移模型的建立 目的建立一種簡便、可行、穩(wěn)定的人結(jié)直腸癌裸小鼠淋巴道移植轉(zhuǎn)移動(dòng)物模型。 方法裸鼠爪墊皮下注射人結(jié)腸癌細(xì)胞系HCT-116細(xì)胞,分別于接種后第1、2、3、4、5、6周及第8周不同時(shí)間采集種植瘤,乆窩和腹股溝淋巴結(jié)(第一級)、髂血管旁淋巴結(jié)(第二級)、腎門淋巴結(jié)(第三級)以及肝、肺臟器標(biāo)本,進(jìn)行HE染色及免疫組化檢測人癌胚抗原(CEA)。觀察種植瘤的成瘤率、生長及淋巴結(jié)轉(zhuǎn)移情況。 結(jié)果爪墊種植1周后成瘤率100%。瘤體在3周后成指數(shù)生長。第一級、第二級、第三級淋巴結(jié)轉(zhuǎn)移率第1、2周均為0%;第3周時(shí)分別為40%、0%、0%;第4周時(shí)轉(zhuǎn)移率分別為60%、0%、0%;第5周時(shí)轉(zhuǎn)移率分別為60%、20%、0%;第6周時(shí)轉(zhuǎn)移率分別為100%、40%、0%。第8周時(shí)轉(zhuǎn)移率分別為100%、60%、20%。但以上各時(shí)點(diǎn)均未見肝、肺轉(zhuǎn)移。HE染色證實(shí)淋巴結(jié)有彌漫性癌細(xì)胞轉(zhuǎn)移浸潤。免疫組化染色顯示轉(zhuǎn)移灶內(nèi)CEA陽性細(xì)胞。 結(jié)論裸鼠爪墊皮下注射人結(jié)腸癌細(xì)胞系(HCT-116)可成功建立人大腸癌淋巴道轉(zhuǎn)移模型。此模型快速簡便、轉(zhuǎn)移集中、轉(zhuǎn)移率高,為研究人大腸癌淋巴道轉(zhuǎn)移機(jī)制,藥物干預(yù)等抗轉(zhuǎn)移治療提供理想的動(dòng)物模型。 第二部分人結(jié)直腸癌裸小鼠直腸粘膜原位種植癌及轉(zhuǎn)移模型的建立 目的建立人大腸癌組織塊裸小鼠直腸粘膜原位種植癌及轉(zhuǎn)移模型,為研究結(jié)直腸癌浸潤轉(zhuǎn)移機(jī)制和抗轉(zhuǎn)移治療提供動(dòng)物模型。 方法人結(jié)腸癌細(xì)胞系HCT-116細(xì)胞注射接種于裸鼠直腸粘膜下,獲得直腸種植瘤。取該腫瘤4周瘤體制備成新鮮組織小塊,原位種植于直腸粘膜面,分別于接種后第2、4、6、8、10、12周不同時(shí)間采集直腸瘤塊,直腸上動(dòng)脈、腸系膜下動(dòng)脈、腹主動(dòng)脈旁淋巴結(jié)以及肝、肺臟器標(biāo)本,進(jìn)行HE染色及人癌胚抗原(CEA)免疫組化檢測。觀察種植瘤的成瘤率,生長及轉(zhuǎn)移情況。 結(jié)果直腸粘膜組織塊法原位種植2周時(shí)成瘤率91%。腫瘤在6~12周成指數(shù)生長。直腸上動(dòng)脈淋巴結(jié)、腸系膜下動(dòng)脈淋巴結(jié)、腹主動(dòng)脈旁淋巴結(jié)轉(zhuǎn)移率第2周時(shí)均為0%,第4周時(shí)分別為60%、20%、0%,第6周時(shí)分別為80%、60%、40%,第8周時(shí)分別為100%、80%、80%,第10、12周時(shí)轉(zhuǎn)移率均為100%。于第10周、12周時(shí)見肝、肺同時(shí)均發(fā)生轉(zhuǎn)移,第10周轉(zhuǎn)移率均為20%;第12周時(shí)肝、肺轉(zhuǎn)移率均為40%。HE染色證實(shí)淋巴結(jié)有彌漫性癌細(xì)胞轉(zhuǎn)移浸潤及肝、肺有轉(zhuǎn)移癌結(jié)節(jié)。免疫組化染色顯示轉(zhuǎn)移灶內(nèi)CEA陽性細(xì)胞。 結(jié)論裸小鼠直腸粘膜植塊法能成功構(gòu)建人結(jié)直腸癌原位種植癌及轉(zhuǎn)移模型。此模型操作簡便、制模穩(wěn)定、轉(zhuǎn)移率高,且生物學(xué)行為更能模擬臨床疾病粘膜原位發(fā)生、發(fā)展演變過程,可為研究大腸癌轉(zhuǎn)移機(jī)制和抗轉(zhuǎn)移治療提供理想的動(dòng)物模型。
[Abstract]:Colorectal cancer is a common tumor in the digestive tract, with high malignancy and easy occurrence of lymphatic metastasis and dissemination of digestive tract. Tumor invasion and metastasis not only affect the effect of clinical treatment, but also the main cause of the recurrence and death of the patients. Therefore, the study on the mechanism of metastasis and the prevention and treatment of colorectal cancer is an important task at present. It is necessary to establish a stable, reliable and reproducible animal model for colorectal cancer transplanting and metastasis in nude mice. It is necessary to establish a stable, reliable and reproducible animal model in order to further study the mechanism of invasion and metastasis of colorectal cancer, drug intervention and other anti metastasis. The treatment provides an effective experimental platform to improve the prognosis and outcome of colorectal cancer. But the ideal model of colorectal carcinoma simple lymphatic metastasis and the model of orthotopic transplantation of colorectal carcinoma have not been established. Therefore, we use the human colon cancer cell line HCT-116 from the model point of view to study the construction of human node. It is of great scientific value and clinical significance that the animal model of simple lymphatic transplanting in the nude mice of rectal cancer and the feasibility of transferring the animal model by the tissue block method of rectal mucosa transplantation in situ are of great importance.
Part one establishment of lymphatic metastasis model of human colorectal cancer in nude mice
Objective to establish a simple, feasible and stable animal model for human lymphatic metastasis of human colorectal cancer in nude mice.
Methods human colon cancer cell line HCT-116 cells were subcutaneously injected into the nude mouse's claw pads. The tumors were collected at the first 1,2,3,4,5,6 and 8 weeks after the inoculation, and the lymph nodes of the socket and groin (grade 1), the iliac para lymph nodes (grade second), the renal hilar lymph nodes (third), and the liver and lung organs were examined by HE staining and immunohistochemical detection. Human carcinoembryonic antigen (CEA). Tumor formation rate, growth and lymph node metastasis were observed.
Results after 1 weeks, the tumor formation rate of the claw pad was grown exponentially after 3 weeks. The first, second, and third lymph node metastasis rates were 0% and 40%, 0%, 0%, respectively, at the third week, and fourth weeks, respectively, 60%, 0%, 0%, respectively. The transfer rate at the week of fourth weeks was respectively, and the metastasis rate of the week was respectively 0%.. The rates were 100%, 60%, 20%., but no liver was found at all time points..HE staining showed that the lymph nodes had diffuse infiltrating infiltration in the lymph nodes. Immunohistochemical staining showed the CEA positive cells in the metastases.
Conclusion the nude mouse claw pad subcutaneous injection of human colon cancer cell line (HCT-116) can successfully establish the lymphatic metastasis model of human colorectal cancer. This model is rapid, simple, centralized and high metastasis rate. It provides an ideal animal model for the study of the mechanism of lymphatic metastasis of colorectal cancer and the antimetastasis treatment of drug intervention.
The second part: establishment of orthotopic implantation cancer and metastasis model of human colorectal cancer in nude mice
Objective to establish the carcinoma and metastasis model of rectal mucosa in the nude mice of colorectal cancer tissue, and to provide an animal model for the study of the mechanism of invasion and metastasis of colorectal cancer and the treatment of anti metastasis.
Methods human colon cancer cell line HCT-116 cells were injected under the rectal mucous membrane of nude mice to obtain rectal implants. The tumor was prepared for 4 weeks by the tumor tissue. The tumor was implanted in the rectal mucosa in situ. The rectal tumor blocks, the superior rectal artery, the inferior mesenteric artery and the abdominal aorta were collected at the different time of the inoculation at the 2,4,6,8,10,12 week. HE staining and human carcinoembryonic antigen (CEA) immunohistochemical staining were used to detect lymph node, liver and lung organ specimens. The tumorigenicity, growth and metastasis of the tumor were observed.
Results the tumor rate of the rectal mucosal tissue mass was in situ for 2 weeks, and the tumor rate 91%. tumor grew exponentially at 6~12 weeks. The upper rectal artery lymph node, the inferior mesenteric artery lymph node, the abdominal aorta lymph node metastasis rate were 0% at second weeks, and 60%, 20%, 0% at the fourth week, respectively, 80%, 60%, 40%, respectively, eighth weeks, respectively, 100%, 80%, 10,1, eighth, respectively. At the 2 week, the metastasis rate was 100%. at tenth weeks, the liver was seen at 12 weeks and the metastasis rate was 20% at the tenth week. The liver and lung metastasis rates were both 40%.HE staining and diffuse infiltration of the lymph nodes and the liver and the metastatic nodules in the lung at the twelfth week. The immunohistochemical staining showed the CEA positive cells in the metastases.
Conclusion the nude mouse rectal mucosal graft method can successfully construct the orthotopic cancer and metastasis model of human colorectal cancer. This model is easy to operate, is stable, and has high metastasis rate. The biological behavior can simulate the occurrence of the clinical disease in situ and the evolution process. It can provide ideal animal for the study of the metastasis mechanism of colorectal cancer and the antimetastasis treatment. Model.

【學(xué)位授予單位】：重慶醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2008
【分類號】：R735.35;R-332

【引證文獻(xiàn)】

相關(guān)碩士學(xué)位論文 前1條

1 黃深;直腸與結(jié)腸注射建立結(jié)直腸癌肝轉(zhuǎn)移動(dòng)物模型安全性與相關(guān)因素分析[D];廣西醫(yī)科大學(xué);2011年

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本文編號：1873764