PDTC对喉癌循环肿瘤细胞的影响及其机制研究

发布时间：2018-06-05 09:47

本文选题：PDTC + 核因子(NF-κB)　；参考：《重庆医科大学》2011年硕士论文

【摘要】：第一部分PDTC对喉癌循环肿瘤细胞的影响目的:利用喉癌裸鼠模型,检测核因子(NF-κB)在喉癌裸鼠模型中的表达情况,了解核因子(NF-κB)与喉癌裸鼠模型中循环肿瘤细胞(circulating tumor cells,CTC)发生的相关性。探讨PDTC对喉癌裸鼠模型中循环肿瘤细胞的作用, 方法:选取4周龄BALB/c-nu裸鼠60只,将其随机分为实验组(20只),对照组(20只)及空白组(20只)。实验组为PDTC组,在接种喉癌细胞后第七天开始按终浓度(100mg/kg)腹腔给药(0.2～0.3ml).对照组为生理盐水组(等量生理盐水),空白组不予接种喉癌细胞。在裸鼠右颈部皮下接种喉癌Hep-2细胞悬液0.2ml。PDTC组及对照组40只裸鼠均成功成瘤。待肿瘤生长8周,处死裸鼠,取出肿瘤组织并取血。肿瘤组织常规HE染色。以CK-19为标志物检测喉癌裸鼠模型中循环肿瘤细胞表达情况,免疫组化法检测核因子(NF-κB)在喉癌组织中的表达,NF-κB激活时由细胞质转到细胞核内,因此以肿瘤细胞核的棕黄色阳性染色反映NF-κB的激活情况。结果: 1.核因子(NF-κB)在喉癌组织中的表达率为65%(26/40)。在CTC表达阳性的喉癌组织中,NF-κB阳性率为90%(9/10),而在CTC表达阴性的喉癌组织中,NF-κB阳性率为56.7%(17/30),通过检验CTC与喉癌组织中NF-κB表达率之间无明显关联。但在NF-κB活性被激活的裸鼠模型中,CTC阳性率为75%(9/12), NF-κB活性未被激活的裸鼠模型中,CTC阳性率为3.5%(1/28)。说明喉癌裸鼠模型中CTC发生与NF-κB是否具有活性有关。 2. PDTC组CTC阳性率5%(1/20) ,对照组CTC阳性率45%(9/20),两组间CTC阳性率有明显差异(P0.005)。结论: 1.喉癌裸鼠模型中,循环肿瘤细胞的发生和核因子(NF-κB)的表达无明显相关,但与核因子(NF-κB)是否具有活性有关。 2. PDTC可抑制喉癌组织中NF-κB的激活,减少喉癌裸鼠模型中CTC的发生率。第二部分PDTC对喉癌循环肿瘤细胞影响的相关机制研究目的:利用喉癌裸鼠模型,观察PDTC使用后,对喉癌组织中VEGF-C和MMP-9mRNA表达的影响,探讨其对喉癌循环肿瘤细胞可能的作用机制,为喉癌的临床治疗提供新思路。方法:选取4周龄BALB/c-nu裸鼠60只,将其随机分为实验组(20只),对照组(20只)及空白组(20只)。实验组为PDTC组,在接种喉癌细胞后第七天开始按终浓度(100mg/kg)腹腔给药(0.2～0.3ml).对照组为生理盐水组(等量生理盐水),空白组不予接种喉癌细胞。在裸鼠右颈部皮下接种喉癌Hep-2细胞悬液0.2ml。PDTC组及对照组40只裸鼠均成功成瘤。待肿瘤生长8周,处死裸鼠,取出肿瘤组织测量其体积并取血。以CK-19为标志物检测裸鼠模型中循环肿瘤细胞表达情况,荧光定量PCR检测各组喉癌组织中VEGF-C和MMP-9mRNA的表达。结果:PDTC组的肿瘤生长受到明显抑制,其体积与对照组差异有统计学意义(P0.01);PDTC组喉癌组织中VEGF-C和MMP-9mRNA的表达较对照组有明显减少。结论:PDTC可抑制喉癌组织中NF-κB的激活,减少喉癌裸鼠模型中CTC的发生率,其作用可能得益PDTC降低了喉癌发生转移的能力,其机制可能为PDTC通过抑制NF-κB活性而使VEGF-C和MMP-9表达下调,减少肿瘤组织新生血管及降低喉癌侵袭力有关。
[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.
【学位授予单位】：重庆医科大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R739.65

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