本文選題：黑色素瘤 + 達(dá)卡巴嗪��； 參考：《華中科技大學(xué)》2010年碩士論文

【摘要】：目的:本研究擬通過動(dòng)物實(shí)驗(yàn)探討IFN-α聯(lián)合治療的可能機(jī)制,以及周細(xì)胞及其新標(biāo)志物RGS5在聯(lián)合治療中可能所起的作用。 方法:將黑色素瘤細(xì)胞株B16細(xì)胞種植在小鼠左前下肢靠背側(cè)皮下,7天后隨機(jī)分成三組:空白對照組,DTIC單藥處理組和DTIC+IFN-α聯(lián)合處理組,每組7只。第16天處死小鼠,測量腫瘤長短徑以計(jì)算腫瘤體積;數(shù)字放射照相術(shù)分析血管形態(tài);免疫組化檢測CD31計(jì)算微血管密度(MVD),檢測α-SMA分析微血管成熟周細(xì)胞覆蓋度和檢測腫瘤HIF-α陽性細(xì)胞的表達(dá)個(gè)數(shù)以分析缺氧區(qū)域;實(shí)時(shí)定量PCR分析RGS5 mRNA的表達(dá);Western Blot檢測RGS5和HIF-α的表達(dá);以及免疫熒光分析RGS5在黑色素瘤中的表達(dá)部位。 結(jié)果:DTIC聯(lián)合IFN-α處理組相對于單用DTIC化療組和對照組,腫瘤生長明顯受到抑制,腫瘤體積顯著下降(對照組5.806±1.436 cm3 vs. DTIC組2.946±1.266 cm3 vs.聯(lián)合治療組1.658±0.949 cm3; P0.05)。IFN-α聯(lián)合處理組還顯示腫瘤血管正�；�,膨大的血管顯著減少,平均血管直徑下降(對照組1.0000±0.5142 mm vs. DTIC組0.8375±0.4676 mm vs.聯(lián)合化療組0.3835±0.2122mm;P0.05);微血管密度(MVD)下降(聯(lián)合化療組21±9 vessels/mm2) vs.對照組62±16 vessels/mm2 vs. DTIC組41±11 vessels/mm2;P0.05);微血管成熟周細(xì)胞覆蓋度顯著上升(DTIC+IFN-α聯(lián)合處理組69.7±16.8% vs. DTIC處理組32±13% vs.對照49±12%;P0.05);腫瘤缺氧顯著緩解(對照組145±13 HIF-α+ cells/field vs. DTIC組119±22 HIF-α+ cells/field vs. IFN-α聯(lián)合化療組66±32 HIF-α+ cells/field;P0.05)。除此之外,我們還發(fā)現(xiàn)黑色素瘤高表達(dá)RGS5蛋白,但是不是表達(dá)在黑色素瘤細(xì)胞,而是表達(dá)在腫瘤管腔周圍。IFN-α聯(lián)合處理組相對于DTIC化療組和對照組,RGS5表達(dá)顯著下降(RGS5 mRNA對照組2.996±0.197 vs. DTIC處理組2.312±0.697 vs.聯(lián)合治療組1.102±0.208, P0.05)。 結(jié)論:本實(shí)驗(yàn)提示黑色素瘤高表達(dá)RGS5蛋白,但是不是表達(dá)在黑色素瘤細(xì)胞,而是表達(dá)在腫瘤管腔周圍——周細(xì)胞。DTIC聯(lián)合IFN-α處理組抗腫瘤效應(yīng)優(yōu)于單用DTIC化療組和對照組部分是由于正�；[瘤血管從而增加傳統(tǒng)化療的效應(yīng)。并且,IFN-α顯示正�；[瘤血管效應(yīng)部分是由于下調(diào)RGS5的表達(dá)及誘導(dǎo)周細(xì)胞成熟,揭示了RGS5是IFN-α的靶點(diǎn)之一,而RGS5特異性表達(dá)細(xì)胞——周細(xì)胞是IFN-α的靶細(xì)胞之一,初步證明了RGS5和周細(xì)胞在聯(lián)合免疫化療治療黑色素瘤中對腫瘤血管正�；心芷鹨欢ǖ淖饔�。
[Abstract]:Aim: to explore the possible mechanism of IFN- 偽 combined therapy and the role of pericytes and its new marker RGS5 in combined therapy by animal experiments. Methods: the melanoma cell line B16 cells were implanted in the subcutaneous side of the left anterior and lower extremity of mice for 7 days, and were randomly divided into three groups: the blank control group treated with DTIC alone and the DTIC IFN- 偽 treated group with 7 rats in each group. On the 16th day, the mice were killed, the tumor length and diameter were measured to calculate the tumor volume, and the vascular morphology was analyzed by digital radiography. CD31 was used to calculate microvessel density (MVD), 偽 -SMA was used to analyze the coverage of microvascular mature pericytes and the number of HIF- 偽 positive cells in tumor to analyze the hypoxia region, the expression of RGS5 mRNA was detected by real-time quantitative PCR and the expression of RGS5 and HIF- 偽 was detected by Western Blot, the expression of RGS5 and HIF- 偽 was detected by Western blot. The expression of RGS5 in melanoma was analyzed by immunofluorescence. Results the tumor growth was significantly inhibited and the tumor volume was significantly decreased (control group 5.806 鹵1.436 cm3 vs 1.436 cm3 vs 2.946 鹵1.266 cm3 vs) in the control group treated with 1: DTIC combined with IFN- 偽. The combined treatment group (1.658 鹵0.949 cm ~ (3); P0.05). IFN- 偽 combined treatment group also showed that the tumor vessels normalized, the enlarged vessels decreased significantly and the mean vessel diameter decreased (control group 1.0000 鹵0.5142 mm vs. DTIC group 0.8375 鹵0.4676 mm vs. The microvessel density (MVD) was decreased (21 鹵9 vessels/mm2) in the combined chemotherapy group (P 0.05). In the control group (62 鹵16 vessels/mm2 vs .DTIC, 41 鹵11 vessels / mm2, P0.05), the coverage of microvascular matured pericytes increased significantly (DTIC IFN- 偽 combined treatment group, 69.7 鹵16.8% vs. DTIC treatment group, 32 鹵13% vs. Tumor anoxia was significantly alleviated in the control group (49 鹵12g / kg) and tumor hypoxia (145 鹵13HIF- 偽 cells/field vs.DTIC group 119 鹵22HIF- 偽 cells/field vs. IFN- 偽 combined chemotherapy group 66 鹵32HIF- 偽 cells / 偽 cells / p0.05). In addition, we also found that melanoma overexpression RGS5 protein, but not in melanoma cells, The expression of RGS5 in the combined treatment group was significantly lower than that in the DTIC chemotherapy group and the control group (RGS5 mRNA was 2.996 鹵0.197 vs 2.312 鹵0.697 vs in the DTIC group). Combined treatment group 1.102 鹵0.208, P0.05). Conclusion: this study suggests that RGS5 protein is highly expressed in melanoma, but not in melanoma cells. But the antitumor effect of the treated group was better than that of the single DTIC chemotherapy group and the control group, partly because of normalizing tumor blood vessels and increasing the effect of traditional chemotherapy. IFN- 偽 showed that the normal vascular effect was partly due to down-regulation of RGS5 expression and induction of pericyte maturation, which revealed that RGS5 was one of the targets of IFN- 偽, and RGS5-specific expression cell-pericyte was one of the target cells of IFN- 偽. It was preliminarily proved that RGS5 and pericytes may play a role in the normalization of tumor blood vessels in combined immunotherapy with melanoma.
【學(xué)位授予單位】：華中科技大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2010
【分類號】：R739.5

【共引文獻(xiàn)】

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本文編號：2113183