本文選題：洛鉑　切入點：射線敏感性　出處：《西南醫(yī)科大學(xué)》2017年碩士論文　論文類型：學(xué)位論文

【摘要】：目的:探討洛鉑抑制宮頸癌SiHa細胞系(cervical cancer cell line SiHa,SiHa)增殖、誘導(dǎo)細胞凋亡及增強細胞射線敏感性的作用及機制。方法:1.以0、1、2.5、5、10、20μmol/L濃度的洛鉑對SiHa細胞的作用24、48和72 h,利用四氮唑溴鹽比色試驗[3-(4,5)-dimethylthiahiazo(-z-y1)-3,5-di-phenytetra-zoliumromide,MTT]檢測洛鉑對宮頸癌SiHa細胞增殖率的影響。2.選取對數(shù)期生長的SiHa細胞接受洛鉑處理,依次接受0、2、4、6、8 Gy劑量的X射線照射48小時,利用單克隆形成實驗檢測洛鉑對SiHa細胞的射線敏感性影響。3.依照處理方式的不同,依次分為空白組、洛鉑組(5μmol/L)、照射組(6 Gy)和洛鉑(5μmol/L)+照射組(6 Gy)。利用流式細胞術(shù)檢測洛鉑、射線以及洛鉑聯(lián)合射線對SiHa細胞凋亡率的影響。4.利用Western blotting檢測洛鉑對促凋亡蛋白Bax和抑凋亡蛋白Bcl-2在SiHa細胞中表達的影響。結(jié)果:1.MTT結(jié)果得出不同濃度的洛鉑處理SiHa細胞24、48和72 h的IC 50值。2.單擊多靶模型擬合單克隆形成實驗數(shù)據(jù)得出空白組和洛鉑處理組的D0值分別為:3.588 Gy,2.226 Gy;空白組和洛鉑組Dq值分別為:0.792 Gy,0.053 Gy;洛鉑在SiHa細胞中的SER值為1.612。3.流式細胞結(jié)果檢測空白組、洛鉑組、射線組和洛鉑聯(lián)合射線組的凋亡率分別為:4.44±2.07%,30.50±3.17%,43.08±5.93%和84.68±4.90%。4.Western blotting檢測發(fā)現(xiàn)洛鉑組、射線組相對空白組促凋亡蛋白Bax表達顯著上調(diào),抑凋亡蛋白Bcl-2顯著下調(diào)。洛鉑聯(lián)合射線組相對洛鉑組、射線組進一步顯著上調(diào)Bax和下調(diào)Bcl-2的表達。結(jié)論:1.洛鉑能夠顯著的抑制宮頸癌SiHa細胞的增殖,隨著洛鉑藥物濃度增大和作用時間的延長SiHa細胞的增殖率逐漸下降。2.洛鉑能顯著增強宮頸癌SiHa細胞的射線敏感性,洛鉑處理后的SiHa細胞在射線下的存活分數(shù)顯著低于未經(jīng)洛鉑處理的SiHa細胞。3.洛鉑和放射照射分別能誘導(dǎo)宮頸癌SiHa細胞凋亡,并且洛鉑聯(lián)合射線能夠更加顯著促進SiHa細胞凋亡。4.洛鉑和放射照射作用于宮頸癌SiHa細胞時,洛鉑和放射照射分別能夠提高促凋亡蛋白Bax的表達水平。洛鉑同步放射照射能提高促凋亡蛋白Bax的表達水平,且顯著高于單獨使用洛鉑或放射照射時。5.洛鉑和放射照射作用于宮頸癌SiHa細胞時,洛鉑和放射照射分別能夠降低抑凋亡蛋白Bcl-2的表達水平。洛鉑同步放射照射能降低抑凋亡蛋白Bcl-2的表達水平,且顯著低單獨使用洛鉑或放射照射時。6.洛鉑通過誘導(dǎo)增加促凋亡蛋白Bax的表達水平、抑制抑凋亡蛋白Bcl-2的表達水平,來實現(xiàn)對宮頸癌SiHa細胞的放療增敏作用。
[Abstract]:Objective: to investigate the inhibitory effect of Loxplatin on the proliferation of cervical carcinoma SiHa cell line, Cervical cancer cell line SiHaSiHa. Effects and mechanisms of inducing apoptosis and enhancing radiosensitivity. Methods the proliferation rate of Loxoplatin on SiHa cells of cervical cancer was detected by MTT assay for 24 h, 48 h and 72 h, respectively. SiHa cells growing in logarithmic phase were treated with loplatin. After 48 hours of X-ray irradiation at a dose of 0, 2, 2, 4, 6 and 8 Gy, the effects of roplatin on the radiosensitivity of SiHa cells were detected by monoclonal formation assay. The cells were divided into blank group according to the different treatment methods. Luoplatin group (5 渭 mol / L) and roplatin (5 渭 mol / L) group (6 Gy) and roplatin (5 渭 mol / L) irradiation group (6 Gy). Flow cytometry was used to detect roplatin. Effects of radiation and Loxplatin combined radiation on apoptosis rate of SiHa cells .4.The effects of Loxplatin on the expression of apoptosis-promoting protein Bax and anti-apoptotic protein Bcl-2 in SiHa cells were detected by Western blotting. Results 1. The results showed that different concentrations of Lopplatin were treated with different concentrations of Lopplatin in SiHa cells. The IC50 value of SiHa cells at 24 h, 48 h and 72 h. Click on the multi-target model to fit the experimental data of monoclonal formation. The D0 values of blank group and loplatin treatment group were: 3.588 Gy / 2.226 Gy; DQ value of blank group and roplatin group were 0. 0.792 Gy / 0. 053 Gy; Lopplatin was in SiHa cells. The SER value in flow cytometry was 1.612.3. The apoptotic rates of Loxoplatin group, radiation group and Loxplatin combined radiation group were 30. 50 鹵3. 17 鹵43. 08 鹵5.93% and 84. 68 鹵4. 90% respectively. Western blotting analysis showed that the expression of apoptotic protein Bax was significantly up-regulated in Lopplatin group, and in the radiation group compared with the control group. The expression of Bax and Bcl-2 were further up-regulated and down-regulated in the radiation group compared with the roplatin group. Conclusion: 1. Lopplatin can significantly inhibit the proliferation of cervical cancer SiHa cells. The proliferation rate of SiHa cells decreased gradually with the increase of Lopplatin concentration and the prolonged time of treatment. Lopplatin could significantly enhance the radiosensitivity of cervical cancer SiHa cells. The survival fraction of SiHa cells treated with Lopplatin was significantly lower than that of SiHa cells without Lopplatin treatment. 3. Loxoplatin and radiation irradiation could induce apoptosis of cervical cancer SiHa cells, respectively. In addition, Loxoplatin combined with radiation can significantly promote apoptosis of SiHa cells. 4. loplatin and radiation irradiation on SiHa cells of cervical cancer. Lopplatin and radiation irradiation increased the expression of apoptosis-promoting protein Bax and the expression of pro-apoptotic protein Bax. And it was significantly higher than that in SiHa cells of cervical cancer treated by Loxplatin alone or radiation irradiation. Lopplatin and radiation could decrease the expression of Bcl-2, and Loxplatin combined with radiation could decrease the expression of Bcl-2. In addition, proapoptotic protein (Bax) expression and inhibition of Bcl-2 expression in cervical cancer SiHa cells were inhibited by low dose of Loxoplatin alone or radiation irradiation, so that the radiosensitization of cervical cancer SiHa cells could be achieved by increasing the expression of apoptosis-promoting protein Bax and inhibiting the expression of apoptosis-inhibiting protein Bcl-2.
【學(xué)位授予單位】：西南醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R96

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