【摘要】：目的:研究白花丹素(Plumbagin,PB)對術(shù)前PF化療方案殺傷舌鱗癌CAL27細(xì)胞凋亡和自噬的影響及作用機(jī)制。方法:1、人舌鱗癌CAL27細(xì)胞的細(xì)胞培養(yǎng)。2、MTT法分別檢測PB、DDP、5-Fu對CAL27細(xì)胞的增殖抑制作用及半數(shù)抑制濃度IC50。3、MTT法檢測PB聯(lián)合PF方案對CAL27細(xì)胞的增殖抑制作用。4、流式細(xì)胞儀檢測PB和/或PF方案作用于舌鱗癌CAL27細(xì)胞后的細(xì)胞凋亡率。5、Cyto ID熒光顯微鏡檢測PB和/或PF方案作用于舌鱗癌CAL27細(xì)胞后的自噬水平。6、Western Blot法檢測PB和/或PF方案作用于舌鱗癌CAL27細(xì)胞后自噬相關(guān)蛋白Beclin1、LC3的表達(dá)。7、Western Blot法檢測PB和/或PF方案作用于舌鱗癌CAL27細(xì)胞后PI3K/AKT/mTOR信號通路蛋白的表達(dá)。結(jié)果:1、PB、DDP、5-Fu分別以濃度依賴方式抑制CAL27細(xì)胞增殖,其作用24h的半數(shù)抑制濃度(IC50)分別為7.77μmol/L、10.50μg/ml、1.26mg/ml。2、PB聯(lián)合PF方案作用,與單獨PF方案比較,舌鱗癌CAL27細(xì)胞的增殖抑制作用明顯增強(qiáng)。3、PB聯(lián)合PF方案作用,與單獨PF方案比較,舌鱗癌CAL27細(xì)胞的凋亡率明顯增強(qiáng)。4、PB聯(lián)合PF方案作用,與單獨PF方案比較,舌鱗癌CAL27細(xì)胞的自噬水平明顯增強(qiáng)。5、當(dāng)PB單獨作用時,CAL27細(xì)胞中PI3K、p-AKT、p-mTOR蛋白表達(dá)不同程度下調(diào),且PB聯(lián)合PF方案比起單獨應(yīng)用PF方案,其細(xì)胞中PI3K、p-AKT、p-mTOR蛋白表達(dá)亦不同程度降低。6、應(yīng)用PI3K/AKT激動劑IGF-1、mTOR激動劑MHY1485作用后,該激動劑逆轉(zhuǎn)了PB對CAL27細(xì)胞中PI3K、p-AKT、p-mTOR蛋白的下調(diào)作用,同時顯著抑制PB對PF化療CAL27細(xì)胞凋亡和自噬誘導(dǎo)作用。結(jié)論:白花丹素通過抑制PI3K/AKT/mTOR信號通路誘導(dǎo)舌鱗癌細(xì)胞凋亡和自噬從而增強(qiáng)PF化療方案殺傷舌鱗癌CAL27細(xì)胞。
[Abstract]:Aim: to study the effect and mechanism of Plumbagin PB on apoptosis and autophagy of CAL27 cells in tongue squamous cell carcinoma treated with PF regimen before operation. Methods the cell culture of human tongue squamous cell carcinoma (CAL27) cells was used to detect the proliferation inhibition of CAL27 cells by PBP 5-Fu, the inhibitory effect of PB combined with PF regimen on the proliferation of CAL27 cells by IC50.3% MTT assay, and the proliferation inhibition of PB by flow cytometry. Apoptosis rate of CAL27 cells in tongue squamous cell carcinoma treated with PF and / or PF. 5 Cyto ID fluorescence microscope to detect autophagy level of PB and / or PF on CAL27 cells of tongue squamous cell carcinoma. Western Blot assay to detect PB and / or PF regimen acting on tongue scale Expression of autophagy associated protein Beclin1hLC3 in CAL27 cells was detected by Western Blot. The expression of PI3K/AKT/mTOR signal pathway protein was detected by PB and / or PF regimen in CAL27 cells of tongue squamous cell carcinoma. Results the proliferation of CAL27 cells was inhibited in a concentration-dependent manner. The half inhibitory concentration (IC50) was 7.77 渭 mol / L (10.50 渭 g / L) and 1.26 mg / ml 路2nPB in combination with PF regimen, respectively. Compared with PF alone, the inhibitory concentration (IC50) was 7.77 渭 mol 路L ~ (-1) 路ml ~ (-1) 路L ~ (-1) 路L ~ (-1) 路L ~ (-1) 路L ~ (-1), respectively. Compared with PF alone, the apoptotic rate of CAL27 cells in tongue squamous cell carcinoma was significantly increased, and compared with PF alone, the apoptotic rate of CAL27 cells in tongue squamous cell carcinoma was significantly increased. The level of autophagy in CAL27 cells of tongue squamous cell carcinoma was significantly increased. When PB was treated alone, the expression of p-mTOR protein was down-regulated in CAL27 cells, and PB combined with PF regimen was significantly lower than that of PF alone. The expression of PI3Knp-AKTor p-mTOR protein was also decreased in different degree. The agonist MHY1485, a PI3K/AKT agonist, reversed the down-regulation of PI3KPT-AKTnp-mTOR protein in CAL27 cells by using IGF-1mTOR agonist. At the same time, PB significantly inhibited apoptosis and autophagy induced by PF chemotherapeutic CAL27 cells. Conclusion: Baihuadan inhibits PI3K/AKT/mTOR signaling pathway and induces apoptosis and autophagy in tongue squamous cell carcinoma cells, thereby enhancing PF chemotherapy regimen to kill CAL27 cells.
【學(xué)位授予單位】：南昌大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R739.86

【參考文獻(xiàn)】

相關(guān)期刊論文 前10條

1 沈杰;許靈敏;王廣磊;;新輔助化療在老年中晚期口腔鱗狀細(xì)胞癌治療中的臨床意義[J];癌癥進(jìn)展;2016年09期

2 劉曉楠;卓姍姍;岳玖玲;張文超;張侖;;新輔助化療在局部晚期口腔癌中的療效分析[J];天津醫(yī)科大學(xué)學(xué)報;2015年05期

3 胡靜;蘇榮健;趙子明;黃克強(qiáng);;自噬介導(dǎo)人舌鱗癌細(xì)胞對順鉑耐藥的作用及其機(jī)制研究[J];醫(yī)學(xué)與哲學(xué)(B);2015年06期

4 宋麗萍;李翠蘭;王子介;湯曦;戴惠如;陳敦金;;自噬與卵巢癌研究進(jìn)展[J];中華臨床醫(yī)師雜志(電子版);2015年03期

5 蔡勁;黎民宇;趙妮妮;施茜;劉金剛;周佳卉;;雷帕霉素聯(lián)合順鉑對舌鱗癌細(xì)胞體外生長影響的研究[J];中國實用醫(yī)刊;2014年20期

6 陳偉泉;譚廣謀;黃文喜;黃海燕;;Beclin-1表達(dá)對舌癌細(xì)胞Tca8113順鉑敏感性的影響[J];山東醫(yī)藥;2014年31期

7 郭海鵬;劉木元;楊熙鴻;彭漢偉;麥瑞琴;;新輔助化療對口腔癌生存率影響的Meta分析[J];汕頭大學(xué)醫(yī)學(xué)院學(xué)報;2014年02期

8 徐璇;潘淑婷;邱嘉旋;;白花丹素增強(qiáng)術(shù)前誘導(dǎo)化療PF方案對舌癌細(xì)胞Tca-8113的殺傷作用[J];實用口腔醫(yī)學(xué)雜志;2014年03期

9 何雙;吳淑華;胡金龍;溫菲菲;孫晨博;;自噬調(diào)控因子beclin1及mTOR在大腸腺瘤癌變過程中的表達(dá)及意義[J];世界華人消化雜志;2014年07期

10 翁軍權(quán);唐海闊;王成;王雅雯;黃秋雨;侯勁松;;沉默Beclin1基因抑制自噬促進(jìn)舌鱗狀細(xì)胞癌細(xì)胞增殖及侵襲轉(zhuǎn)移[J];中華口腔醫(yī)學(xué)研究雜志(電子版);2013年01期

，

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