常見人體腫瘤轉(zhuǎn)移動物模型的建立及相關(guān)機理研究
發(fā)布時間：2018-08-29 13:07
【摘要】： 轉(zhuǎn)移性是惡性腫瘤主要的生物學(xué)特性之一，也是導(dǎo)致腫瘤患者死亡的主要原因，即使對患者實施腫瘤切除，術(shù)后轉(zhuǎn)移的發(fā)生也是影響手術(shù)療效的主要障礙。腫瘤轉(zhuǎn)移的發(fā)生發(fā)展是一個多因素、多步驟的復(fù)雜生物學(xué)過程。因此，研究腫瘤的轉(zhuǎn)移，就必然要建立與臨床表現(xiàn)相近的腫瘤轉(zhuǎn)移動物模型。人體腫瘤移植于裸小鼠建立人癌異種移植動物模型為腫瘤的深入研究提供了可能。皮下移植和原位移植是目前常用的兩種方法。原位移植由于獲得與人體腫瘤生長相似的微環(huán)境，更利于腫瘤惡性行為的表達，因此，建立腫瘤轉(zhuǎn)移動物模型大多采用原位移植的方法。肝癌、胃癌和肺癌是常見的腫瘤，利用這三種腫瘤進行研究具有一定的代表意義。本課題利用不同程度免疫缺陷小鼠觀察了人肝癌原位移植腫瘤在宿主體內(nèi)的生物學(xué)特性差異；模擬臨床，建立了人肝癌和人肺癌術(shù)后轉(zhuǎn)移動物模型；同時利用經(jīng)體內(nèi)篩選建立的人胃癌高轉(zhuǎn)移動物模型以及建立的人胃癌細胞株MKN-45mc，對腫瘤轉(zhuǎn)移的相關(guān)機制進行探討。 一、目的 1．建立人肝癌NOD-SCID小鼠和裸小鼠原位移植模型，并通過兩種模型生物學(xué)特性的比較，對不同類型免疫缺陷動物在人肝癌異種移植方面的應(yīng)用價值進行初步探討。 2．建立人肝癌術(shù)后轉(zhuǎn)移動物模型，觀察并比較手術(shù)切除腫瘤后對人肝癌小鼠皮下移植模型和原位移植模型的影響。 3．建立裸小鼠人肺癌術(shù)后轉(zhuǎn)移模型，為研究肺癌的轉(zhuǎn)移機制和術(shù)后抗轉(zhuǎn)移治療提供適宜的動物模型。 4．比較體內(nèi)篩選前后建立的人胃癌原位移植動物模型，以及人胃癌細胞株MKN-45mc和MKN-45生物學(xué)特性出現(xiàn)的差異，對轉(zhuǎn)移的相關(guān)機制進行探討。 二、方法 1．將組織結(jié)構(gòu)完整的SMMC-LTNM瘤塊植入NOD-SCID小鼠和裸小鼠肝臟內(nèi)，建立人肝癌原位移植模型。觀察成瘤率、腫瘤體積和重量、臟器的轉(zhuǎn)移情況以及血清甲胎蛋白(AFP)、γ-谷氨酰轉(zhuǎn)肽酶同工酶Ⅱ(γ-GTⅡ)等指標(biāo)。 2．將組織結(jié)構(gòu)完整的HC-031瘤塊植入NOD-SCID小鼠皮下和肝臟內(nèi)，分別建立人肝癌皮下移植和原位移植模型。模擬臨床腫瘤手術(shù)切除方法，切除皮下移植瘤和原位移植瘤，建立肝癌術(shù)后轉(zhuǎn)移模型。觀察腫瘤生長和動物的生存狀況。通過病理解剖和組織病理學(xué)技術(shù)研究腫瘤在動物體內(nèi)轉(zhuǎn)移情況。 3．將人非小細胞肺癌NCI-H460組織塊植入裸小鼠皮下，建立皮下移植瘤模型。4w后，25只裸小鼠作為手術(shù)組切除腫瘤組織，建立術(shù)后轉(zhuǎn)移模型，15只裸小鼠作為對照組。每兩周兩組各處死5只動物動態(tài)觀察其遠處各臟器轉(zhuǎn)移情況。動物明顯消瘦時結(jié)束實驗。采用免疫組化方法檢測MMP-2、OPN、CD44v6、CD62等蛋白在轉(zhuǎn)移灶及原發(fā)灶的表達情況。 4．用人胃癌MKN-45sci和MKN-45腫瘤組織塊建立人胃癌原位移植動物模型，觀察并比較兩種模型的腫瘤生長和轉(zhuǎn)移、血清腫瘤標(biāo)志物及MMP-2、OPN、CD44v6、CD62與Timp-2的表達情況。取MKN-45sci肝轉(zhuǎn)移灶形成的皮下移植瘤，用組織塊法進行原代培養(yǎng)，建立MKN-45mc細胞株，觀察腫瘤細胞的形態(tài)學(xué)、生長速度、體外侵襲能力、細胞周期變化，并與MKN-45細胞株進行比較。 三、結(jié)果 1．NOD-SCID小鼠人肝癌原位移植模型于5w可捫及腫瘤的生長，成瘤率為100％，11w腫瘤體積為(4.48±0.93)cm~3，瘤重為(7.02±1.15)g，肺部轉(zhuǎn)移率為53.85％；裸小鼠人肝癌原位移植模型于6w～7w可捫及腫瘤的生長，成瘤率為100％，11w腫瘤體積為(1.02±0.70)cm~3，瘤重為(2.87±0.44)g，體內(nèi)未見轉(zhuǎn)移發(fā)生。NOD-SCID小鼠的瘤體積、瘤重、肺轉(zhuǎn)移率均高于裸小鼠(P=0.000，P=0.000，P=0.011)。二者均保持AFP高分泌和γ-GT同工酶陽性的特性。 2．人肝癌HC-031皮下移植模型和原位移植模型由于腫瘤負荷過大分別于11w和6w而出現(xiàn)瀕死狀態(tài)，荷瘤平均生存時間為75d和44d。病理解剖，皮下移植瘤模型未發(fā)現(xiàn)轉(zhuǎn)移(0／7)，原位移植瘤模型的腫瘤和鄰近臟器侵襲明顯，部分出現(xiàn)肺部轉(zhuǎn)移(2／7)。皮下切瘤動物和肝原位切瘤動物分別于17w和12w出現(xiàn)惡病質(zhì)現(xiàn)象，平均生存時間為154d和112d。病理解剖發(fā)現(xiàn)肺部均有肉眼可見癌性轉(zhuǎn)移結(jié)節(jié)(7／7，7／7)。未切瘤動物和切瘤動物血清中AFP和γ-GTⅡ表達均為陽性。 3．人肺癌NCI-H460裸小鼠皮下移植瘤10w時瘤體嚴重壞死，并且動物出現(xiàn)明顯消瘦，解剖未發(fā)現(xiàn)轉(zhuǎn)移。手術(shù)組裸小鼠于10w、12w各發(fā)現(xiàn)一例肺轉(zhuǎn)移(1／5)，14w時裸小鼠明顯消瘦，肺轉(zhuǎn)移率達100％(5／5)，有肉眼明顯可見的肺轉(zhuǎn)移結(jié)節(jié)，其他臟器未見轉(zhuǎn)移。免疫組化結(jié)果表明，MMP-2、OPN、CD44v6、CD62和Timp-2在肺轉(zhuǎn)移灶內(nèi)的表達明顯高于原發(fā)灶，CD54和MMP-9在轉(zhuǎn)移灶內(nèi)的表達則低于原發(fā)灶。 4．人胃癌原位移植模型MKN-45sci組動物的肝轉(zhuǎn)移出現(xiàn)早且轉(zhuǎn)移率高，4w左右肉眼即可看到肝轉(zhuǎn)移結(jié)節(jié)(100％)，同時伴有淋巴結(jié)轉(zhuǎn)移(100％)、肺轉(zhuǎn)移(71％)、脾轉(zhuǎn)移(29％)和腹水；移植瘤體積和重量明顯增加，分別達到(3089±1617)mm~3和(2.66±1.32)g，動物出現(xiàn)惡病質(zhì)。而4w時MKN-45組肝、肺、脾未見明顯轉(zhuǎn)移灶，僅2只動物有淋巴結(jié)轉(zhuǎn)移；小鼠未見明顯消瘦，移植瘤體積和重量為(275±90)mm~3和(0.35±0.14)g。MKN-45sci組血清中NSE和CYFRA21-1的濃度均較高，且隨著腫瘤生長時間的延長而增高。而MKN-45組CYFRA21-1始終呈現(xiàn)陰性，4w時NSE僅為76.9 ng／ml。免疫組化結(jié)果顯示MMP-9、OPN抗體在MKN-45sci組原位移植瘤和轉(zhuǎn)移灶呈強陽性反應(yīng)，而在MKN-45組原位移植瘤反應(yīng)較弱。CD44v6抗體在MKN-45sci組原位移植瘤和轉(zhuǎn)移灶呈陽性反應(yīng)。E-cadherin抗體在MKN-45sci組原位移植瘤呈陽性反應(yīng)，肝轉(zhuǎn)移灶呈陰性反應(yīng)。建立的MKN-45mc細胞株的染色體為超三倍體細胞，具有人類惡性腫瘤細胞染色體的特點。細胞形態(tài)為典型的上皮樣細胞，與MKN-45的形態(tài)學(xué)特點相似。流式細胞分析顯示MKN-45mc與MKN-45細胞周期各時相比例分別為：G_0-G_1期62.51％／53.95％，S期9.46％／1452％，G_2-M期28.04％／31.53％，兩株細胞DNA合成期的細胞比例均較高。MKN-45mc與MKN-45細胞倍增時間分別為34.8h和42.1h，前者的生長速度高于后者。MKN-45mc與MKN-45的36h體外過膜數(shù)分別為(60.38±8.86)／高倍視野和(32.50±17.26)／高倍視野，前者多于后者。 四、結(jié)論 1．建立了與臨床表現(xiàn)相似的人肝癌動物模型。與裸小鼠相比，NOD-SCID小鼠在建立人肝癌的異種移植模型方面有更大的應(yīng)用價值。 2．模擬臨床腫瘤切除方法，建立人肝癌術(shù)后轉(zhuǎn)移動物模型。實驗結(jié)果表明，荷瘤動物腫瘤切除后，生存期延長，有利于腫瘤轉(zhuǎn)移發(fā)生。 3．裸小鼠人肺癌術(shù)后轉(zhuǎn)移模型模擬了臨床腫瘤根除術(shù)后發(fā)生遠處轉(zhuǎn)移的過程，，結(jié)果提示肺癌轉(zhuǎn)移的發(fā)生可能與部分粘附分子的異常表達相關(guān)，同時為研究肺癌轉(zhuǎn)移機制和術(shù)后抗轉(zhuǎn)移治療提供理想的動物模型。 4．人胃癌原位移植動物模型體內(nèi)篩選前后腫瘤的生物學(xué)特性不同，經(jīng)篩選后腫瘤的生長和轉(zhuǎn)移能力較未篩選的增強，其機理與腫瘤細胞粘附和降解能力得到強化有關(guān)。篩選后建立的細胞株生長速度、過膜能力比原細胞株增強，表明腫瘤轉(zhuǎn)移與腫瘤細胞增殖和運動變形能力有關(guān)。
[Abstract]:Metastasis is one of the main biological characteristics of malignant tumors, and also the main cause of death of cancer patients. Even after tumor resection, the occurrence of postoperative metastasis is also a major obstacle to the efficacy of surgery. It is necessary to establish an animal model of tumor metastasis which is similar to the clinical manifestation. Human tumor transplantation in nude mice provides the possibility of establishing an animal model of human cancer xenotransplantation. Liver cancer, gastric cancer and lung cancer are common tumors, and the use of these three kinds of tumors has a certain representative significance. In this study, immunodeficiency mice were used to observe the orthotopic transplantation of human liver cancer in vivo. The animal models of human hepatocellular carcinoma and human lung cancer metastasis were established by simulating clinical conditions, and the high metastasis animal models of human gastric cancer and the human gastric cancer cell line MKN-45mc were established by screening in vivo.
First, the purpose
1. The orthotopic transplantation models of human hepatocellular carcinoma NOD-SCID mice and nude mice were established, and the application value of different immunodeficient animals in xenotransplantation of human hepatocellular carcinoma was preliminarily discussed by comparing the biological characteristics of the two models.
2. Establish the animal model of human liver cancer metastasis after operation, observe and compare the effect of tumor resection on subcutaneous transplantation model and orthotopic transplantation model of human liver cancer mice.
3. To establish a metastasis model of human lung cancer in nude mice, and provide an appropriate animal model for studying the metastasis mechanism and anti-metastasis therapy.
4. To compare the animal models of orthotopic transplantation of human gastric cancer established before and after screening in vivo and the differences of biological characteristics between human gastric cancer cell lines MKN-45mc and MKN-45, and to explore the mechanism of metastasis.
Two, method
1. SMMC-LTNM tumor with intact tissue was implanted into the liver of NOD-SCID mice and nude mice to establish orthotopic transplantation model of human hepatocellular carcinoma.
2. HC-031 tumor was implanted into NOD-SCID mice subcutaneously and in the liver to establish the models of subcutaneous transplantation and original transplantation of human hepatocellular carcinoma. Anatomical and histopathological techniques were used to study the metastasis of tumors in animals.
3. Human non-small cell lung cancer (NSCLC) NCI-H460 was implanted subcutaneously into nude mice to establish a subcutaneous transplanted tumor model. Four weeks later, 25 nude mice were used as surgical group to remove tumor tissues, and 15 nude mice as control group. Five animals in each group were killed every two weeks to observe the distant organ metastasis. Immunohistochemistry was used to detect the expression of MMP-2, OPN, CD44v6 and CD62 in metastatic and primary lesions.
4. The orthotopic transplantation model of human gastric cancer was established by using MKN-45sci and MKN-45 tumor tissue blocks. The growth and metastasis of tumor, the expression of serum tumor markers and MMP-2, OPN, CD44v6, CD62 and Timp-2 were observed and compared between the two models. MKN-45 MC cell line was established to observe the morphology, growth rate, invasion ability in vitro and cell cycle changes of tumor cells, and compared with MKN-45 cell line.
Three, the result
1. Human hepatocellular carcinoma orthotopic transplantation model in NOD-SCID mice could palpate the growth of tumor in 5 weeks, the tumor formation rate was 100%, the tumor volume was (4.48.93) cm~3, the tumor weight was (7.02 1.15) g, and the lung metastasis rate was 53.85%; human hepatocellular carcinoma orthotopic transplantation model in nude mice could palpate the growth of tumor in 6 to 7 weeks, the tumor formation rate was 100%, and the tumor volume was (1.02.7) at 11 weeks. The tumor volume, tumor weight and lung metastasis rate of NOD-SCID mice were higher than those of nude mice (P = 0.000, P = 0.000, P = 0.011). Both of them maintained the characteristics of high AFP secretion and positive gamma-GT isozyme.
2. The subcutaneous transplantation model and the orthotopic transplantation model of HC-031 were in the state of near death due to the overload of the tumor at 11th and 6th weeks, respectively. The average survival time of the transplanted tumor was 75 and 44 days. Pathological anatomy showed that no metastasis was found in the subcutaneous transplantation model (0/7). The tumor and adjacent organs of the orthotopic transplantation model had obvious invasion and partial lung metastasis (2). The average survival time was 154 D and 112 D in subcutaneous and in situ tumor-resected animals, respectively. The metastatic nodules (7/7, 7/7) were found in the lung by pathological dissection. The expression of AFP and gamma-GT II was positive in the serum of both non-resected and resected animals.
3. Human lung cancer NCI-H460 nude mice underwent subcutaneous transplantation of tumor for 10 weeks, and the tumor necrosis was severe, and the animals showed significant emaciation and no metastasis. One case of lung metastasis (1/5) was found in each of the nude mice in the operation group at 10 and 12 weeks, and the metastasis rate was 100% (5/5) in the nude mice at 14 weeks, with obvious metastasis nodules in the lung and no metastasis in other organs. Metastasis. Immunohistochemistry showed that the expressions of MMP-2, OPN, CD44v6, CD62 and Timp-2 were significantly higher in lung metastases than in primary lesions, while the expressions of CD54 and MMP-9 were lower in metastases.
4. Liver metastasis was early and high in MKN-45sci group, with lymph node metastasis (100%), lung metastasis (71%), spleen metastasis (29%) and ascites. The volume and weight of transplanted tumor increased significantly, reaching (3089+1617) mm~3 and (2.66+1.32) g, respectively. Cachexia was found in the mice, but no metastasis was found in the liver, lung and spleen of MKN-45 group at 4 weeks, only 2 animals had lymph node metastasis, and the mice did not show obvious emaciation. The serum NSE and CYFRA21-1 concentrations in MKN-45 SCI group were higher than those in MKN-45 group with (275+90) mm~3 and (0.35+0.14) G. The results of immunohistochemistry showed that MMP-9, OPN antibody and MKN-45sci were strongly positive in transplanted tumors and metastases, but weakly in MKN-45. CD44v6 antibody was positive in transplanted tumors and metastases in MKN-45sci. The established MKN-45mc cell line is a Hypertriploid cell with the characteristics of human malignant tumor cell chromosomes. The cell morphology is typical epithelioid cells, similar to the morphological characteristics of MKN-45. Flow cytometry analysis showed that MKN-45mc and MKN-45 cell cycle. The proportions of the two cell lines were 62.51%/53.95% in G_0-G_1 phase, 9.46%/1452% in S phase and 28.04%/31.53% in G_2-M phase, respectively. The doubling time of MKN-45mc and MKN-45 cells were 34.8 h and 42.1 h, respectively. The growth rate of the former was higher than that of the latter. The number of outer membrane of MKN-45mc and MKN-45 cells in 36 h was 60.46 h, respectively. 38 + 8.86) / high field of view and (32.50 + 17.26) / high power field of vision, the former is more than the latter.
Four. Conclusion
1. Establishment of human hepatocellular carcinoma animal model with similar clinical manifestations. Compared with nude mice, NOD-SCID mice have more application value in establishing xenograft model of human hepatocellular carcinoma.
2. To simulate the clinical tumor resection method and establish the animal model of human liver cancer metastasis after operation. The experimental results show that the survival time of tumor-bearing animals after tumor resection is prolonged, which is conducive to the occurrence of tumor metastasis.
3. The metastasis model of human lung cancer in nude mice simulates the process of distant metastasis after clinical tumor eradication. The results suggest that the metastasis of lung cancer may be related to the abnormal expression of some adhesion molecules, and provide an ideal animal model for studying the metastasis mechanism of lung cancer and anti-metastasis therapy.
4. The biological characteristics of human gastric cancer orthotopic transplantation animal models were different before and after screening in vivo. The growth and metastasis ability of the tumor after screening was stronger than that of the non-screening. The mechanism was related to the enhancement of adhesion and degradation ability of tumor cells. Tumor metastasis is related to cell proliferation and movement deformability.
【學(xué)位授予單位】：南方醫(yī)科大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2007
【分類號】：R-332;R730.2
本文編號：2211300
[Abstract]:Metastasis is one of the main biological characteristics of malignant tumors, and also the main cause of death of cancer patients. Even after tumor resection, the occurrence of postoperative metastasis is also a major obstacle to the efficacy of surgery. It is necessary to establish an animal model of tumor metastasis which is similar to the clinical manifestation. Human tumor transplantation in nude mice provides the possibility of establishing an animal model of human cancer xenotransplantation. Liver cancer, gastric cancer and lung cancer are common tumors, and the use of these three kinds of tumors has a certain representative significance. In this study, immunodeficiency mice were used to observe the orthotopic transplantation of human liver cancer in vivo. The animal models of human hepatocellular carcinoma and human lung cancer metastasis were established by simulating clinical conditions, and the high metastasis animal models of human gastric cancer and the human gastric cancer cell line MKN-45mc were established by screening in vivo.
First, the purpose
1. The orthotopic transplantation models of human hepatocellular carcinoma NOD-SCID mice and nude mice were established, and the application value of different immunodeficient animals in xenotransplantation of human hepatocellular carcinoma was preliminarily discussed by comparing the biological characteristics of the two models.
2. Establish the animal model of human liver cancer metastasis after operation, observe and compare the effect of tumor resection on subcutaneous transplantation model and orthotopic transplantation model of human liver cancer mice.
3. To establish a metastasis model of human lung cancer in nude mice, and provide an appropriate animal model for studying the metastasis mechanism and anti-metastasis therapy.
4. To compare the animal models of orthotopic transplantation of human gastric cancer established before and after screening in vivo and the differences of biological characteristics between human gastric cancer cell lines MKN-45mc and MKN-45, and to explore the mechanism of metastasis.
Two, method
1. SMMC-LTNM tumor with intact tissue was implanted into the liver of NOD-SCID mice and nude mice to establish orthotopic transplantation model of human hepatocellular carcinoma.
2. HC-031 tumor was implanted into NOD-SCID mice subcutaneously and in the liver to establish the models of subcutaneous transplantation and original transplantation of human hepatocellular carcinoma. Anatomical and histopathological techniques were used to study the metastasis of tumors in animals.
3. Human non-small cell lung cancer (NSCLC) NCI-H460 was implanted subcutaneously into nude mice to establish a subcutaneous transplanted tumor model. Four weeks later, 25 nude mice were used as surgical group to remove tumor tissues, and 15 nude mice as control group. Five animals in each group were killed every two weeks to observe the distant organ metastasis. Immunohistochemistry was used to detect the expression of MMP-2, OPN, CD44v6 and CD62 in metastatic and primary lesions.
4. The orthotopic transplantation model of human gastric cancer was established by using MKN-45sci and MKN-45 tumor tissue blocks. The growth and metastasis of tumor, the expression of serum tumor markers and MMP-2, OPN, CD44v6, CD62 and Timp-2 were observed and compared between the two models. MKN-45 MC cell line was established to observe the morphology, growth rate, invasion ability in vitro and cell cycle changes of tumor cells, and compared with MKN-45 cell line.
Three, the result
1. Human hepatocellular carcinoma orthotopic transplantation model in NOD-SCID mice could palpate the growth of tumor in 5 weeks, the tumor formation rate was 100%, the tumor volume was (4.48.93) cm~3, the tumor weight was (7.02 1.15) g, and the lung metastasis rate was 53.85%; human hepatocellular carcinoma orthotopic transplantation model in nude mice could palpate the growth of tumor in 6 to 7 weeks, the tumor formation rate was 100%, and the tumor volume was (1.02.7) at 11 weeks. The tumor volume, tumor weight and lung metastasis rate of NOD-SCID mice were higher than those of nude mice (P = 0.000, P = 0.000, P = 0.011). Both of them maintained the characteristics of high AFP secretion and positive gamma-GT isozyme.
2. The subcutaneous transplantation model and the orthotopic transplantation model of HC-031 were in the state of near death due to the overload of the tumor at 11th and 6th weeks, respectively. The average survival time of the transplanted tumor was 75 and 44 days. Pathological anatomy showed that no metastasis was found in the subcutaneous transplantation model (0/7). The tumor and adjacent organs of the orthotopic transplantation model had obvious invasion and partial lung metastasis (2). The average survival time was 154 D and 112 D in subcutaneous and in situ tumor-resected animals, respectively. The metastatic nodules (7/7, 7/7) were found in the lung by pathological dissection. The expression of AFP and gamma-GT II was positive in the serum of both non-resected and resected animals.
3. Human lung cancer NCI-H460 nude mice underwent subcutaneous transplantation of tumor for 10 weeks, and the tumor necrosis was severe, and the animals showed significant emaciation and no metastasis. One case of lung metastasis (1/5) was found in each of the nude mice in the operation group at 10 and 12 weeks, and the metastasis rate was 100% (5/5) in the nude mice at 14 weeks, with obvious metastasis nodules in the lung and no metastasis in other organs. Metastasis. Immunohistochemistry showed that the expressions of MMP-2, OPN, CD44v6, CD62 and Timp-2 were significantly higher in lung metastases than in primary lesions, while the expressions of CD54 and MMP-9 were lower in metastases.
4. Liver metastasis was early and high in MKN-45sci group, with lymph node metastasis (100%), lung metastasis (71%), spleen metastasis (29%) and ascites. The volume and weight of transplanted tumor increased significantly, reaching (3089+1617) mm~3 and (2.66+1.32) g, respectively. Cachexia was found in the mice, but no metastasis was found in the liver, lung and spleen of MKN-45 group at 4 weeks, only 2 animals had lymph node metastasis, and the mice did not show obvious emaciation. The serum NSE and CYFRA21-1 concentrations in MKN-45 SCI group were higher than those in MKN-45 group with (275+90) mm~3 and (0.35+0.14) G. The results of immunohistochemistry showed that MMP-9, OPN antibody and MKN-45sci were strongly positive in transplanted tumors and metastases, but weakly in MKN-45. CD44v6 antibody was positive in transplanted tumors and metastases in MKN-45sci. The established MKN-45mc cell line is a Hypertriploid cell with the characteristics of human malignant tumor cell chromosomes. The cell morphology is typical epithelioid cells, similar to the morphological characteristics of MKN-45. Flow cytometry analysis showed that MKN-45mc and MKN-45 cell cycle. The proportions of the two cell lines were 62.51%/53.95% in G_0-G_1 phase, 9.46%/1452% in S phase and 28.04%/31.53% in G_2-M phase, respectively. The doubling time of MKN-45mc and MKN-45 cells were 34.8 h and 42.1 h, respectively. The growth rate of the former was higher than that of the latter. The number of outer membrane of MKN-45mc and MKN-45 cells in 36 h was 60.46 h, respectively. 38 + 8.86) / high field of view and (32.50 + 17.26) / high power field of vision, the former is more than the latter.
Four. Conclusion
1. Establishment of human hepatocellular carcinoma animal model with similar clinical manifestations. Compared with nude mice, NOD-SCID mice have more application value in establishing xenograft model of human hepatocellular carcinoma.
2. To simulate the clinical tumor resection method and establish the animal model of human liver cancer metastasis after operation. The experimental results show that the survival time of tumor-bearing animals after tumor resection is prolonged, which is conducive to the occurrence of tumor metastasis.
3. The metastasis model of human lung cancer in nude mice simulates the process of distant metastasis after clinical tumor eradication. The results suggest that the metastasis of lung cancer may be related to the abnormal expression of some adhesion molecules, and provide an ideal animal model for studying the metastasis mechanism of lung cancer and anti-metastasis therapy.
4. The biological characteristics of human gastric cancer orthotopic transplantation animal models were different before and after screening in vivo. The growth and metastasis ability of the tumor after screening was stronger than that of the non-screening. The mechanism was related to the enhancement of adhesion and degradation ability of tumor cells. Tumor metastasis is related to cell proliferation and movement deformability.
【學(xué)位授予單位】：南方醫(yī)科大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2007
【分類號】：R-332;R730.2
【引證文獻】
相關(guān)期刊論文 前1條
1 錢曉玲;郭勇;;丹參酮ⅡA對腫瘤轉(zhuǎn)移影響的實驗展望[J];深圳中西醫(yī)結(jié)合雜志;2010年06期
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