【摘要】：目的:研究阿霉素(ADM)與二甲雙胍(MET)單獨(dú)應(yīng)用及二者聯(lián)合用藥對(duì)體外培養(yǎng)的舌癌CAL27細(xì)胞增殖以及凋亡的影響。方法:1.依次采用MET藥物濃度5mmol/L、10mmol/L、20mmol/L、40mmol/L、80mmol/L、160mmol/L。ADM藥物濃度0.02mg/L、0.04mg/L、0.08mg/L、0.16mg/L、0.32mg/L。分別對(duì)體外培養(yǎng)的舌癌CAL27細(xì)胞作用24h、48h、72h后,用CCK-8法檢測(cè)各用藥組的吸光度值,算出細(xì)胞的抑制率,利用SPSS 17.0軟件計(jì)算出兩組藥物單獨(dú)使用時(shí)抑制50%細(xì)胞生長(zhǎng)的濃度IC50;而聯(lián)合用藥組則采取單獨(dú)用藥時(shí)所得的ADM組IC50+MET組IC50處理CAL27細(xì)胞48h后上酶標(biāo)儀檢測(cè)吸光度值,計(jì)算出聯(lián)合用藥組的細(xì)胞抑制率;2.根據(jù)CCK-8實(shí)驗(yàn)組數(shù)據(jù),選取48h MET半抑制濃度10mmol/L,ADM半抑制濃度0.05mg/L及二者聯(lián)合作用于CAL27細(xì)胞后,用流式細(xì)胞術(shù)檢測(cè)各實(shí)驗(yàn)組細(xì)胞凋亡的情況;3.采用q RT-PCR技術(shù)檢測(cè)MET 10mmol/L,ADM 0.05mg/L及MET 10mmol/L+ADM 0.05mg/L聯(lián)合應(yīng)用48h后舌癌CAL27細(xì)胞的相關(guān)凋亡基因Bax及Bcl-2 m RNA相對(duì)表達(dá)的情況。結(jié)果:1.CCK-8數(shù)據(jù)顯示MET與ADM單獨(dú)作用24h、48h、72h后,均對(duì)舌癌CAL27細(xì)胞的增殖產(chǎn)生抑制作用,并且在一定濃度范圍內(nèi)呈劑量與時(shí)間的依賴性,二者聯(lián)合應(yīng)用時(shí)比單獨(dú)用藥對(duì)CAL27細(xì)胞的增殖抑制作用更加明顯。2.流式細(xì)胞術(shù)結(jié)果顯示經(jīng)MET 10mmol/L+ADM 0.05mg/L聯(lián)合用藥后比單獨(dú)藥物組更能促進(jìn)細(xì)胞的凋亡。3.q RT-PCR顯示MET及ADM二者合用時(shí)舌癌CAL27細(xì)胞的抗細(xì)胞凋亡基因Bcl-2的相對(duì)表達(dá)水平與單獨(dú)用藥組相比下調(diào)趨勢(shì)更明顯,而促細(xì)胞凋亡基因Bax m RNA的相對(duì)表達(dá)與單獨(dú)用藥組相比上調(diào)趨勢(shì)更明顯。結(jié)論:二甲雙胍作為降血糖藥物與阿霉素作用相似,單獨(dú)使用時(shí)具有對(duì)舌癌CAL27細(xì)胞體外增殖的抑制作用。當(dāng)二者聯(lián)合應(yīng)用時(shí),對(duì)舌癌CAL27細(xì)胞的增殖抑制及促進(jìn)凋亡的效果比單獨(dú)用藥更為顯著。
[Abstract]:Aim: to study the effects of adriamycin (ADM) and metformin (MET) on the proliferation and apoptosis of tongue cancer CAL27 cells cultured in vitro. Methods: 1. The drug concentration of MET was 5 mmol / L 10 mmol / L and 20 mmol / L = 40 mmol / L = 80 mmol / L = 160 mmol / L = 0.02 mg / L 0.04 mg / L 0.08 mg / L = 0.16 mg / L = 0.32 mg / L. After cultured tongue cancer CAL27 cells were treated for 24 h or 48 h for 72 h, the absorbance of each drug group was measured by CCK-8 method, and the inhibition rate of the cells was calculated. SPSS 17.0 software was used to calculate the concentration of IC50; that inhibited 50% cell growth in two groups of drugs alone. However, in the combined treatment group, the CAL27 cells in the IC50 MET group were treated with IC50 for 48 hours, and the absorbance value was measured by enzyme labeling instrument, and the cell inhibition rate of the combined treatment group was calculated. 2. According to the data of CCK-8 experimental group, 10 mmol / L MET semi-inhibitory concentration (0.05mg/L) of 48 h and their combination were used to detect the apoptosis of CAL27 cells by flow cytometry. 3. Q RT-PCR technique was used to detect the relative expression of Bax and Bcl-2 m RNA in CAL27 cells of tongue cancer treated with MET 10 mmol / L 0.05mg/L and MET 10mmol/L ADM 0.05mg/L for 48 h. Results: 1.CCK-8 data showed that the proliferation of tongue cancer CAL27 cells was inhibited by MET and ADM alone for 24 h or 48 h or 72 h, and in a dose-dependent and time-dependent manner. The inhibitory effect of the two drugs on the proliferation of CAL27 cells was more obvious than that of the drug alone. 2. 2. The results of flow cytometry showed that MET 10mmol/L ADM 0.05mg/L combined with drugs could promote cell apoptosis more than that of single drug group. 3. Q RT-PCR showed antiapoptotic genes in CAL27 cells of tongue cancer combined with MET and ADM. The relative expression level of Bcl-2 was more down-regulated than that of the control group. The relative expression of apoptosis-promoting gene Bax m RNA was significantly higher than that of the control group. Conclusion: metformin, as a hypoglycemic drug, can inhibit the proliferation of tongue cancer CAL27 cells in vitro. When combined, the effect of inhibiting proliferation and promoting apoptosis in CAL27 cells of tongue cancer was more significant than that of drug alone.
【學(xué)位授予單位】：廣西醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R739.86

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