發(fā)布時間：2018-06-05 09:47

  本文選題：PDTC + 核因子(NF-κB)��； 參考：《重慶醫(yī)科大學(xué)》2011年碩士論文

【摘要】：第一部分PDTC對喉癌循環(huán)腫瘤細胞的影響 目的:利用喉癌裸鼠模型,檢測核因子(NF-κB)在喉癌裸鼠模型中的表達情況,了解核因子(NF-κB)與喉癌裸鼠模型中循環(huán)腫瘤細胞(circulating tumor cells,CTC)發(fā)生的相關(guān)性。探討PDTC對喉癌裸鼠模型中循環(huán)腫瘤細胞的作用, 方法:選取4周齡BALB/c-nu裸鼠60只,將其隨機分為實驗組(20只),對照組(20只)及空白組(20只)。實驗組為PDTC組,在接種喉癌細胞后第七天開始按終濃度(100mg/kg)腹腔給藥(0.2～0.3ml).對照組為生理鹽水組(等量生理鹽水),空白組不予接種喉癌細胞。在裸鼠右頸部皮下接種喉癌Hep-2細胞懸液0.2ml。PDTC組及對照組40只裸鼠均成功成瘤。待腫瘤生長8周,處死裸鼠,取出腫瘤組織并取血。腫瘤組織常規(guī)HE染色。以CK-19為標(biāo)志物檢測喉癌裸鼠模型中循環(huán)腫瘤細胞表達情況,免疫組化法檢測核因子(NF-κB)在喉癌組織中的表達,NF-κB激活時由細胞質(zhì)轉(zhuǎn)到細胞核內(nèi),因此以腫瘤細胞核的棕黃色陽性染色反映NF-κB的激活情況。 結(jié)果: 1.核因子(NF-κB)在喉癌組織中的表達率為65%(26/40)。在CTC表達陽性的喉癌組織中,NF-κB陽性率為90%(9/10),而在CTC表達陰性的喉癌組織中,NF-κB陽性率為56.7%(17/30),通過檢驗CTC與喉癌組織中NF-κB表達率之間無明顯關(guān)聯(lián)。但在NF-κB活性被激活的裸鼠模型中,CTC陽性率為75%(9/12), NF-κB活性未被激活的裸鼠模型中,CTC陽性率為3.5%(1/28)。說明喉癌裸鼠模型中CTC發(fā)生與NF-κB是否具有活性有關(guān)。 2. PDTC組CTC陽性率5%(1/20) ,對照組CTC陽性率45%(9/20),兩組間CTC陽性率有明顯差異(P0.005)。 結(jié)論: 1.喉癌裸鼠模型中,循環(huán)腫瘤細胞的發(fā)生和核因子(NF-κB)的表達無明顯相關(guān),但與核因子(NF-κB)是否具有活性有關(guān)。 2. PDTC可抑制喉癌組織中NF-κB的激活,減少喉癌裸鼠模型中CTC的發(fā)生率。第二部分PDTC對喉癌循環(huán)腫瘤細胞影響的相關(guān)機制研究 目的:利用喉癌裸鼠模型,觀察PDTC使用后,對喉癌組織中VEGF-C和MMP-9mRNA表達的影響,探討其對喉癌循環(huán)腫瘤細胞可能的作用機制,為喉癌的臨床治療提供新思路。 方法:選取4周齡BALB/c-nu裸鼠60只,將其隨機分為實驗組(20只),對照組(20只)及空白組(20只)。實驗組為PDTC組,在接種喉癌細胞后第七天開始按終濃度(100mg/kg)腹腔給藥(0.2～0.3ml).對照組為生理鹽水組(等量生理鹽水),空白組不予接種喉癌細胞。在裸鼠右頸部皮下接種喉癌Hep-2細胞懸液0.2ml。PDTC組及對照組40只裸鼠均成功成瘤。待腫瘤生長8周,處死裸鼠,取出腫瘤組織測量其體積并取血。以CK-19為標(biāo)志物檢測裸鼠模型中循環(huán)腫瘤細胞表達情況,熒光定量PCR檢測各組喉癌組織中VEGF-C和MMP-9mRNA的表達。 結(jié)果:PDTC組的腫瘤生長受到明顯抑制,其體積與對照組差異有統(tǒng)計學(xué)意義(P0.01);PDTC組喉癌組織中VEGF-C和MMP-9mRNA的表達較對照組有明顯減少。 結(jié)論:PDTC可抑制喉癌組織中NF-κB的激活,減少喉癌裸鼠模型中CTC的發(fā)生率,其作用可能得益PDTC降低了喉癌發(fā)生轉(zhuǎn)移的能力,其機制可能為PDTC通過抑制NF-κB活性而使VEGF-C和MMP-9表達下調(diào),減少腫瘤組織新生血管及降低喉癌侵襲力有關(guān)。
[Abstract]:Part 1 Effect of PDTC on circulating tumor cells in laryngeal carcinoma
Objective: to detect the expression of nuclear factor (NF- kappa B) in the nude mouse model of laryngeal carcinoma by using the nude mouse model of larynx cancer and to understand the correlation between nuclear factor (NF- kappa B) and the occurrence of circulating tumor cells (circulating tumor cells, CTC) in the nude mouse model of larynx cancer and to explore the effect of PDTC on the circulating tumor cells in the nude mouse model of laryngeal carcinoma.
Methods: 60 BALB/c-nu nude mice of 4 weeks old were randomly divided into experimental group (20 rats), control group (20 rats) and blank group (20 rats). The experimental group was group PDTC, and the final concentration (0.2 to 0.3ml) was administered at the final concentration (0.2 to 0.3ml) after the inoculation of larynx cancer cells. The control group was a saline group (equal amount of saline), and the blank group was not inoculated with larynx cancer. Cells in the right neck of the nude mice were subcutaneously inoculated with the Hep-2 cell suspension 0.2ml.PDTC of the larynx cancer cell suspension and the control group of 40 nude mice. After 8 weeks of tumor growth, the tumor tissues were killed and the blood was taken out. The tumor tissue was stained with the conventional HE staining. The expression of the circulating tumor cells in the nude mouse model of larynx cancer was detected with CK-19 as a marker, and the immuno histochemical method was used to detect the nucleus. The expression of factor (NF- kappa B) in the carcinoma of the larynx, and the activation of NF- kappa B from the cytoplasm to the nucleus. Therefore, the activation of NF- kappa B is reflected by the brown yellow positive staining of the nucleus of the tumor.
Result:
The expression rate of 1. nuclear factor (NF- kappa B) in the larynx tissues was 65% (26/40). The positive rate of NF- kappa B was 90% (9/10) in the CTC positive larynx tissues, but the positive rate of NF- kappa B was 56.7% (17/30) in the negative CTC of the larynx tissues. In nude mice, the positive rate of CTC was 75% (9/12), and the positive rate of CTC was 3.5% (1/28) in the nude mice model of NF- kappa B activity, which indicated that the occurrence of CTC in the mouse model of larynx cancer was related to the activity of NF- kappa B.
2. the positive rate of CTC in group PDTC was 5% (1/20), and the positive rate of CTC in control group was 45% (9/20). There was a significant difference in the positive rate of CTC between the two groups (P0.005).
Conclusion:
1. in the nude mice model of laryngeal carcinoma, the occurrence of circulating tumor cells was not related to the expression of nuclear factor (NF- kappa B), but it was related to the activity of nuclear factor NF- (B).
2. PDTC can inhibit the activation of NF- kappa B in laryngeal carcinoma and reduce the incidence of CTC in the nude mouse model of larynx cancer. The study of the related mechanism of the effect of second part PDTC on the tumor cells of laryngeal carcinoma
Objective: To observe the effect of PDTC on the expression of VEGF-C and MMP-9mRNA in laryngeal carcinoma and to explore the possible mechanism of its action on the circulating tumor cells of larynx cancer by using the nude mouse model of larynx cancer, and to provide a new idea for the clinical treatment of larynx cancer.
Methods: 60 BALB/c-nu nude mice of 4 weeks old were randomly divided into experimental group (20 rats), control group (20 rats) and blank group (20 rats). The experimental group was group PDTC, and the final concentration (0.2 to 0.3ml) was administered at the final concentration (0.2 to 0.3ml) after the inoculation of larynx cancer cells. The control group was a saline group (equal amount of saline), and the blank group was not inoculated with larynx cancer. Cells in the right neck of the nude mice were subcutaneously inoculated with Hep-2 cell suspension 0.2ml.PDTC of larynx cancer cell suspension and 40 nude mice in the control group. The tumor tissue was killed for 8 weeks. The tumor tissue was taken out to measure its volume and take blood. The expression of circulating tumor cells in the nude mouse model was detected with CK-19 as a marker, and the fluorescence quantitative PCR was used to detect the larynx tissues in each group. The expression of VEGF-C and MMP-9mRNA.
Results: the tumor growth in the PDTC group was significantly inhibited, and the difference between the volume and the control group was statistically significant (P0.01), and the expression of VEGF-C and MMP-9mRNA in the PDTC group was significantly lower than that in the control group.
Conclusion: PDTC can inhibit the activation of NF- kappa B in laryngeal carcinoma and reduce the incidence of CTC in the nude mouse model of larynx cancer. The effect of PDTC may benefit from the decrease of the ability of PDTC to metastasize. The mechanism may be that PDTC can reduce the expression of VEGF-C and MMP-9 through the inhibition of NF- kappa B activity, reduce the neovascularization of tumor tissue and reduce the invasiveness of larynx cancer.
【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】：2011
【分類號】：R739.65

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本文編號：1981557