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  本文選題：鼻咽癌　切入點：中藥　出處：《廣西醫(yī)科大學(xué)》2010年碩士論文

【摘要】： 背景和目的鼻咽癌是我國南方常見的惡性腫瘤之一,目前以放、化療治療鼻咽癌,只能使5年生存率達到50%～70%,鼻咽癌放化療耐受和治療并發(fā)癥嚴(yán)重影響治療效果。尋找對鼻咽癌治療更有效、毒副反應(yīng)更小的藥物,提高療效是臨床治療迫切需要解決的問題。傳統(tǒng)中醫(yī)藥是人們在生活、疾病中不斷總結(jié)、篩選而得,經(jīng)過臨床實踐和基礎(chǔ)研究發(fā)現(xiàn)多種中草藥有抗腫瘤作用,但是否有抗鼻咽癌作用仍有待研究。本課題采用體外抗鼻咽癌中藥篩選模型,比較5種廣西常見中藥的抗鼻咽癌效果,篩選出一種較佳的抗鼻咽癌藥物,并對其抗鼻咽癌作用機制進行研究。 方法通過提取中藥、血清藥理學(xué)、體外MTT實驗方法,比較白花蛇舌草、半枝蓮、仙鶴草、石上柏、兩面針,5種中藥提取物和含藥血清對人鼻咽癌CNE-1、CNE-2細胞生長的抑制率。用體外抗鼻咽癌作用較強的石上柏提取物,觀察對小鼠的急性毒性,處理鼻咽癌細胞和正常人胚肺成纖維細胞株HFL-I后,MTT法檢測抑制率,倒置相差顯微鏡觀察細胞的形態(tài)學(xué)改變,碘化丙啶(PI)標(biāo)記流式細胞術(shù)檢測鼻咽癌細胞周期分布,Annexin V-FITC和PI標(biāo)記流式細胞術(shù)檢測細胞凋亡,免疫細胞化學(xué)檢測凋亡相關(guān)蛋白Bax、Bcl-2的表達變化。 結(jié)果石上柏的提取物和含藥血清對CNE1、CNE2細胞生長均有明顯抑制作用,有劑量-效應(yīng)和時間-效應(yīng)關(guān)系;高濃度的兩面針提取物和其低濃度含藥血清對CNE1、CNE2細胞生長有一定抑制作用;較高濃度的白花蛇舌草提取物、半枝蓮提取物和仙鶴草提取物對CNE1、CNE2生長有一定的抑制作用,呈劑量-效應(yīng)和時間-效應(yīng)關(guān)系,但其含藥血清對CNE1、CNE2生長未見抑制作用。 2.5mg/mL石上柏提取物可顯著抑制鼻咽癌細胞的生長,而對正常人胚肺成纖維細胞株HFL-I細胞生長影響較小;小鼠經(jīng)720g/kg石上柏提取物灌胃,7天內(nèi)無死亡,肝腎功能和病理切片未見明顯異常;石上柏提取物作用于鼻咽癌細胞后,細胞生長受抑,形態(tài)改變;流式細胞術(shù)檢測細胞周期分布顯示,石上柏提取物誘導(dǎo)鼻咽癌細胞S期阻滯;流式細胞術(shù)檢測細胞凋亡顯示,鼻咽癌細胞凋亡率隨著石上柏提取物濃度的升高而逐漸升高;免疫細胞化學(xué)實驗顯示,石上柏提取物處理后的細胞與對照組細胞相比:Bax蛋白表達增強,Bcl-2蛋白表達減少。 結(jié)論石上柏提取物和含藥血清在體外能顯著抑制人鼻咽癌細胞生長,體內(nèi)和體外急性毒性較小;誘導(dǎo)鼻咽癌細胞S期阻滯,可能為細胞周期特異性藥物,誘導(dǎo)鼻咽癌細胞凋亡,其作用機制與Bax、Bcl-2蛋白表達改變有關(guān)。
[Abstract]:Background and objective Nasopharyngeal carcinoma (NPC) is one of the most common malignant tumors in southern China. The 5-year survival rate can only reach 50% and 70%. The radiotherapy and chemotherapy tolerance and treatment complications of nasopharyngeal carcinoma (NPC) have a serious impact on the effectiveness of the treatment. Look for drugs that are more effective and less toxic in the treatment of nasopharyngeal carcinoma. To improve the curative effect is an urgent problem to be solved in clinical treatment. Traditional Chinese medicine is a kind of traditional Chinese medicine that has been continuously summarized and screened in people's lives and diseases. Through clinical practice and basic research, it has been found that many kinds of Chinese herbal medicines have anti-tumor effects. However, whether it has anti-nasopharyngeal carcinoma effect is still to be studied. In this paper, we compared the anti-nasopharyngeal carcinoma effect of five kinds of common traditional Chinese medicine in Guangxi by using the screening model of anti-nasopharyngeal medicine in vitro, and screened out a better anti-nasopharyngeal cancer drug. The mechanism of its anti-nasopharyngeal carcinoma was studied. Methods by extracting traditional Chinese medicine, serum pharmacology and MTT method in vitro, the comparison was made between Scutellaria chinensis, Scutellaria barbata, Herba Scutellariae and Cypress sclerotii. The inhibitory rate of five kinds of traditional Chinese medicine extracts and serum containing drugs on the growth of human nasopharyngeal carcinoma (NPC) CNE-1 CNE-2 cells was studied. The acute toxicity of the extracts of Cypress sclerotifolia, which has a strong effect on nasopharyngeal carcinoma, was observed in vitro. After HFL-I was treated with nasopharyngeal carcinoma cells and normal human embryonic lung fibroblasts, the inhibition rate was detected by MTT assay, and the morphological changes of the cells were observed by inverted phase contrast microscope. Annexin V-FITC and Pi labeled flow cytometry were used to detect apoptosis and immunocytochemistry to detect the expression of apoptosis-related protein Bcl-2. Results the extracts and serum of Phellodendron lanceolata significantly inhibited the growth of CNE 2 cells in a dose-effect and time-response relationship. The growth of CNE1CNE2 cells was inhibited by high concentration of Acanthopsis nitifolia extract and its low concentration serum, and the growth of CNE1CNE2 cells was inhibited by higher concentration extract of Herba Scutellariae, Herba Scutellariae and Herba Herba Sinensis. There was a dose-effect and time-effect relationship, but the growth of CNE _ 2 was not inhibited by its serum. The growth of nasopharyngeal carcinoma (NPC) cells was significantly inhibited by the extract of 2.5mg/mL, but had little effect on the growth of normal human embryonic lung fibroblast cell line HFL-I, and no death was found in mice fed with 720g/kg extract. There were no obvious abnormalities in liver and kidney function and pathological sections. The growth of nasopharyngeal carcinoma cells was inhibited and the morphology of nasopharyngeal carcinoma cells was changed after treated with the extract of Phellodendron sclerotifolia. Flow cytometry was used to detect the distribution of cell cycle. The apoptosis of nasopharyngeal carcinoma cells was detected by flow cytometry. The apoptosis rate of nasopharyngeal carcinoma cells increased with the increase of extract concentration. The immunocytochemical experiment showed that the apoptosis rate of nasopharyngeal carcinoma cells increased with the increase of extract concentration. Compared with the control group, the expression of Bcl 2 protein was significantly increased in the cells treated with the extract of Cypress sclerotii, and the expression of Bcl 2 protein was decreased. Conclusion the extracts of Phellodendron lanceolata and serum containing drugs can significantly inhibit the growth of human nasopharyngeal carcinoma cells in vitro and have little acute toxicity in vivo and in vitro, and induce S phase arrest of nasopharyngeal carcinoma cells, which may be cell cycle specific drugs. The mechanism of inducing apoptosis of nasopharyngeal carcinoma cells is related to the change of Baxon Bcl-2 protein expression.
【學(xué)位授予單位】：廣西醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2010
【分類號】：R739.63

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