本文選題：表皮生長(zhǎng)因子酪氨酸激酶抑制劑 + 非小細(xì)胞肺癌��； 參考：《第二軍醫(yī)大學(xué)》2016年博士論文

【摘要】：研究背景:表皮生長(zhǎng)因子酪氨酸激酶抑制劑(EGFR-TKI)已經(jīng)成為EGFR基因突變的晚期非小細(xì)胞肺癌(NSCLC)的標(biāo)準(zhǔn)治療之一,然而EGFR-TKI的獲得性耐藥已成為影響其療效的瓶頸。目前化療仍是EGFR-TKI耐藥后的最重要治療手段,然而化療療效的預(yù)測(cè)和個(gè)體化至今難以實(shí)現(xiàn),如何選擇更有效的化療方案意義重大。鑒于既往研究顯示:EGFR-TKI獲得性耐藥后對(duì)不同化療藥物敏感性不同,經(jīng)典的T790M突變和非經(jīng)典的上皮間質(zhì)轉(zhuǎn)換(EMT)等機(jī)制與二線化療療效相關(guān),我們首次提出科學(xué)假設(shè):EGFR-TKI的不同耐藥機(jī)制可能影響后續(xù)化療療效。研究方法:通過EGFR-TKI誘導(dǎo)肺癌細(xì)胞PC-9(EGFR突變型)和H460(EGFR野生型)獲得經(jīng)典的T790M突變的細(xì)胞株P(guān)C-9/ZD,EMT陽(yáng)性的耐藥細(xì)胞株P(guān)C-9/GR和H460/ER,CCK8法檢測(cè)不同肺癌細(xì)胞株對(duì)化療藥物的敏感性;通過RT-PCR、Western-blot檢測(cè)EGFR-TKI獲得性耐藥機(jī)制;si RNA干擾下調(diào)T790M突變細(xì)胞的T790M基因、轉(zhuǎn)染CDH1后逆轉(zhuǎn)耐藥細(xì)胞EMT,檢測(cè)各細(xì)胞對(duì)化療藥物敏感性變化闡明T790M和EMT對(duì)化療敏感性的影響;裸鼠成瘤實(shí)驗(yàn)進(jìn)一步驗(yàn)證化療療效;裸鼠肺轉(zhuǎn)移模型檢測(cè)T790M突變和EMT陽(yáng)性對(duì)裸鼠肺轉(zhuǎn)移、生存的影響。研究結(jié)果:1、對(duì)目前治療NSCLC的第三代化療藥物(吉西他濱、培美曲塞、紫杉醇、多西他塞、伊力替康),EGFR野生型肺癌細(xì)胞株H460的敏感性差于EGFR基因突變的細(xì)胞株P(guān)C-9;而對(duì)順鉑的敏感性兩者無(wú)差異。2、EGFR-TKI獲得性耐藥后的PC-9/ZD細(xì)胞(T790M突變,EMT陰性)與親代PC-9細(xì)胞相比,對(duì)化療藥物(紫杉醇、多西他塞)的敏感增高,但對(duì)化療藥物(順鉑、吉西他濱、培美曲塞)的藥物敏感性無(wú)差異。PC-9/ZD(T790M突變,EMT陰性)對(duì)化療藥物敏感性(紫杉醇、多西他塞)優(yōu)于PC-9/GR(無(wú)T790M突變,EMT陽(yáng)性)。3、特異性下調(diào)EGFR和T790M突變基因后,NSCLC細(xì)胞株P(guān)C-9/ZD對(duì)化療藥物(紫杉醇、多西他塞)敏感性均下降。4、EMT陽(yáng)性NSCLC細(xì)胞株(PC-9/GR和H460/ER)對(duì)化療藥物(順鉑、吉西他濱、培美曲塞、紫杉醇、多西他塞)敏感性差于親代PC-9和H460細(xì)胞株;TGF-β1作用肺癌細(xì)胞PC-9和H460后對(duì)上述化療藥物敏感性下降;PC-9/GR和H460/ER細(xì)胞轉(zhuǎn)染CDH1后逆轉(zhuǎn)EMT,可以恢復(fù)細(xì)胞對(duì)上述化療藥物的敏感性。5、EGFR-TKI獲得性耐藥細(xì)胞株,EMT陽(yáng)性肺癌細(xì)胞(PC-9/GR)裸鼠肺轉(zhuǎn)移數(shù)目多于T790M突變細(xì)胞PC-9/ZD,進(jìn)一步影響裸鼠的生存(EMT陽(yáng)性PC-9/GR細(xì)胞肺轉(zhuǎn)移裸鼠生存期短于T790M突變肺轉(zhuǎn)移裸鼠)。結(jié)論:EGFR-TKI獲得性耐藥后細(xì)胞不同耐藥機(jī)制(T790M突變和EMT陽(yáng)性)影響后續(xù)的化療療效,T790M是紫杉類化療敏感性的指標(biāo),EMT與化療耐藥有關(guān);T790M突變、EMT陽(yáng)性還是遠(yuǎn)處轉(zhuǎn)移和預(yù)后的指標(biāo),有T790M突變的肺癌細(xì)胞株肺轉(zhuǎn)移數(shù)目少,預(yù)后好;而EMT陽(yáng)性肺癌細(xì)胞株肺轉(zhuǎn)移數(shù)目多,生存期短。
[Abstract]:Background: EGFR-TKI, an inhibitor of epidermal growth factor tyrosine kinase, has become one of the standard treatments for advanced non-small cell lung cancer (NSCLC) with EGFR gene mutation. However, the acquired resistance of EGFR-TKI has become a bottleneck affecting its efficacy. At present, chemotherapy is still the most important treatment method after EGFR-TKI resistance. However, it is difficult to predict and individualize the effect of chemotherapy, so how to choose a more effective chemotherapy regimen is of great significance. Since previous studies have shown that the sensitivity to different chemotherapeutic agents is different after acquired drug resistance, the mechanisms of classical T790M mutation and non-classical epithelial interstitial transition (EMTT) are related to the efficacy of second-line chemotherapy. For the first time, we suggest that different drug resistance mechanisms of 1: EGFR-TKI may affect the efficacy of subsequent chemotherapy. Methods: the chemosensitivity of different lung cancer cell lines to chemotherapeutic drugs was detected by EGFR-TKI induced PC-9 EGFR mutation) and H460 EGFR wild-type). The classical T790M mutant cell lines PC-9 / ZDN-EMT positive drug resistant cell lines PC-9% gr and H460% ERCCK8 were used to detect the chemosensitivity of different lung cancer cell lines to chemotherapeutic drugs. EGFR-TKI acquired drug resistance mechanism (EGFR-TKI) was detected by RT-PCR Western-blot. The T790M gene was down-regulated by siRNA interference in EGFR-TKI mutant cells. After transfection of CDH1, the EMTs were reversed. The effects of T790M and EMT on the chemosensitivity of EGFR-TKI cells were investigated. T790M mutation and EMT positive effect on lung metastasis and survival of nude mice were detected in nude mice lung metastasis model. The third generation of chemotherapy drugs (gemcitabine, pemetrexed, paclitaxel, docetaxel, docetaxel) for current treatment of NSCLC, The sensitivity of wild-type lung cancer cell line H460 with EGFR gene mutation was lower than that of PC-9 cell line with EGFR gene mutation, but the sensitivity to cisplatin was not different between H460 cell line and PC-9 cell line, but the sensitivity to cisplatin was not different between H460 cell line and PC-9 cell line, but the sensitivity to cisplatin was not different between H460 cell line and PC-9 cell line. Sensitivity to chemotherapeutic drugs (paclitaxel, docetaxel) was increased, but there was no difference in sensitivity to chemotherapeutic drugs (cisplatin, gemcitabine, pemetrexide). PC-9 / ZDT790M mutation EMT was negative for chemotherapeutic drugs (paclitaxel, paclitaxel). Docetaxel was superior to PC-9 / GR( no T790M mutation EMT positive, down-regulated the sensitivity of EGFR and T790M mutant PC-9 / ZD to chemotherapeutic drugs (paclitaxel, docetaxel). 4% EMT positive NSCLC cell lines PC-9% GR and H460% ERR) were more sensitive to chemotherapeutic drugs (cisplatin, docetaxel). Gemcitabine, pemetrexed, paclitaxel, The sensitivity of PC-9 and H460 cells to these chemotherapeutic drugs decreased after transfection of PC-9 / gr and H460 / ER cells, and then reversed EMTs, which could restore the sensitivity of the cells to the above chemotherapeutic drugs after transfection of PC-9 and H460 cell lines with TGF- 尾 1. The number of lung metastasis in nude mice was more than that in T790M mutant PC-9 / ZD, which further affected the survival of Nude mice with EMT positive PC-9 / gr cells and the survival time of Nude mice with EMT positive PC-9 / gr cells was shorter than that of T790M mutant lung metastasis nude mice. Conclusion the T790M mutation and EMT positive mechanism of different resistance mechanisms after acquired drug resistance in the cells of 10% EGFR-TKI affect the therapeutic effect of subsequent chemotherapy. T790M is an indicator of the chemosensitivity of Taxus chinensis. The relationship between EMT and chemotherapeutic resistance is related to the EMT of T790M mutation or distant metastasis and prognosis. Lung cancer cell lines with T790M mutation had fewer metastatic numbers and better prognosis, while EMT positive lung cancer cell lines had more metastatic numbers and shorter survival time.
【學(xué)位授予單位】：第二軍醫(yī)大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2016
【分類號(hào)】：R734.2

【相似文獻(xiàn)】

相關(guān)期刊論文 前10條

1 陸智祥,沈曉梅,楊同華;老年人急性非淋巴細(xì)胞白血病化療療效觀察[J];新醫(yī)學(xué);2000年01期

2 孫明東,楊勝,鄒霓,李尊昌;老年急性髓性白血病21例化療療效觀察[J];臨床薈萃;2001年24期

3 羅文紀(jì) ,劉興貴 ,桑桂珍 ,史軍;70歲以上老年急性髓細(xì)胞白血病化療療效觀察[J];臨床醫(yī)學(xué);2002年12期

4 ;智能“化療炸彈”有望提高化療療效[J];技術(shù)與市場(chǎng);2011年05期

5 孫貴富,黃光,趙忠仁;中晚期非小細(xì)胞肺癌年齡因素對(duì)化療療效的影響[J];中國(guó)腫瘤臨床與康復(fù);2001年02期

6 王立富,陳夕志,王書祥,耿素萍;晚期肺癌支氣管動(dòng)脈灌注化療療效分析[J];江蘇臨床醫(yī)學(xué)雜志;1999年06期

7 顧康生;腫瘤的基因表達(dá)與化療療效的預(yù)測(cè)[J];癌癥;2001年12期

8 張連生，，施鑫鶴，顧莉莉;初治怠性非淋巴細(xì)胞白血病P─170的表達(dá)與化療療效的關(guān)系[J];甘肅科技;1996年02期

9 王紹娟;魏萍;李家梅;梁勁荃;;卵巢癌基因表達(dá)與化療療效及預(yù)后相關(guān)性的研究[J];臨床和實(shí)驗(yàn)醫(yī)學(xué)雜志;2006年10期

10 寧芳;何曉蓮;才紹江;;影響兒童急性淋巴細(xì)胞白血病化療療效及預(yù)后的分子生物學(xué)因素[J];中國(guó)小兒血液與腫瘤雜志;2009年05期

相關(guān)會(huì)議論文 前10條

1 顧雅佳;馮曉源;邱龍華;彭衛(wèi)軍;楊文濤;費(fèi)菲;陳燦敏;;磁共振彌散成像對(duì)局部進(jìn)展期乳腺癌輔助化療療效評(píng)價(jià)的初步研究[A];中華醫(yī)學(xué)會(huì)第十三屆全國(guó)放射學(xué)大會(huì)論文匯編（下冊(cè)）[C];2006年

2 王丹;吳鳴;任彤;萬(wàn)希潤(rùn);馮鳳芝;黃惠芳;楊佳欣;沈鏗;向陽(yáng);;化療前血紅蛋白及血小板水平對(duì)子宮頸癌新輔助化療后手術(shù)患者化療療效及預(yù)后的影響[A];中華醫(yī)學(xué)會(huì)第十次全國(guó)婦產(chǎn)科學(xué)術(shù)會(huì)議婦科腫瘤會(huì)場(chǎng)（婦科腫瘤學(xué)組、婦科病理學(xué)組）論文匯編[C];2012年

3 楊傳敏;;正確看待中藥治療癌癥的作用[A];2011年全國(guó)醫(yī)藥學(xué)術(shù)論壇交流會(huì)暨臨床藥學(xué)與藥學(xué)服務(wù)研究進(jìn)展培訓(xùn)班論文集[C];2011年

4 詹文華;閆鋼;折虹;;62例Ⅲ期非小細(xì)胞肺癌三維適形放射治療聯(lián)合化療療效分析[A];中華醫(yī)學(xué)會(huì)放射腫瘤治療學(xué)分會(huì)六屆二次暨中國(guó)抗癌協(xié)會(huì)腫瘤放療專業(yè)委員會(huì)二屆二次學(xué)術(shù)會(huì)議論文集[C];2009年

5 張錦林;倪美鑫;劉繼斌;沈康;張一心;;補(bǔ)康靈對(duì)Lewis肺癌小鼠化療療效及免疫功能的影響[A];2010年江蘇省藥學(xué)大會(huì)暨第十屆江蘇省藥師周大會(huì)論文集[C];2010年

6 黨彩玲;陽(yáng)志軍;李力;;復(fù)發(fā)上皮卵巢癌化療療效的Meta分析[A];中華醫(yī)學(xué)會(huì)第十次全國(guó)婦產(chǎn)科學(xué)術(shù)會(huì)議婦科腫瘤會(huì)場(chǎng)（婦科腫瘤學(xué)組、婦科病理學(xué)組）論文匯編[C];2012年

7 李力;莫秀瑛;張瑋;韋敏怡;;多基因蛋白表達(dá)聯(lián)合檢查判斷卵巢癌化療療效的初步探討[A];中華醫(yī)學(xué)會(huì)第九次全國(guó)婦科腫瘤學(xué)術(shù)會(huì)議論文匯編[C];2006年

8 周宇靈;章文成;陳俠斌;肖澤芬;喬艷;于典科;林東昕;譚文;;轉(zhuǎn)化生長(zhǎng)因子-β1基因多態(tài)與食管鱗癌放療和化療療效及生存的關(guān)系[A];全國(guó)腫瘤流行病學(xué)和腫瘤病因?qū)W學(xué)術(shù)會(huì)議論文集[C];2011年

9 張超;崔恒;李藝;祝洪瀾;梁旭東;李小平;魏麗惠;沈丹華;虞有智;陳定寶;;原發(fā)性腹膜惡性腫瘤的免疫組化研究及其預(yù)后相關(guān)分析[A];中華醫(yī)學(xué)會(huì)第九次全國(guó)婦科腫瘤學(xué)術(shù)會(huì)議論文匯編[C];2006年

10 閻飛;張定昌;朱愛琴;;血清中NSE水平對(duì)晚期NSCLC化療療效的影響[A];第四屆中國(guó)腫瘤學(xué)術(shù)大會(huì)暨第五屆海峽兩岸腫瘤學(xué)術(shù)會(huì)議論文集[C];2006年

相關(guān)重要報(bào)紙文章 前1條

1 張中橋;中國(guó)~(18)F—FDG顯像評(píng)價(jià)化療療效[N];醫(yī)藥經(jīng)濟(jì)報(bào);2007年

相關(guān)博士學(xué)位論文 前2條

1 喬曉娟;1.肺癌耐藥相關(guān)基因在化療療效評(píng)估的基礎(chǔ)和臨床研究 2.晚期肺腺癌患者EGFR-TKI治療耐藥后治療策略的臨床分析[D];北京市結(jié)核病胸部腫瘤研究所;2015年

2 周娟;T790M及EMT在預(yù)測(cè)EGFR-TKI繼發(fā)耐藥后化療療效中的重要作用及機(jī)制[D];第二軍醫(yī)大學(xué);2016年

相關(guān)碩士學(xué)位論文 前10條

1 陳秋彬;基于超聲RF信號(hào)頻譜分析的腫瘤化療療效早期評(píng)價(jià)[D];華南理工大學(xué);2015年

2 李政華;老年晚期NSCLC患者ERCC1、RRM1、TUBB3的表達(dá)與化療療效及預(yù)后的相關(guān)性研究[D];陜西中醫(yī)藥大學(xué);2015年

3 劉培培;非小細(xì)胞肺癌患者血清CEA變化率與化療療效及預(yù)后的相關(guān)性[D];鄭州大學(xué);2014年

4 沈志勇;肺癌合并胸腔積液患者化療療效的臨床指標(biāo)的觀察[D];福建醫(yī)科大學(xué);2013年

5 張明慧;晚期非小細(xì)胞肺癌化療療效分析及化療敏感性預(yù)測(cè)[D];第三軍醫(yī)大學(xué);2009年

6 曲顏麗;肺癌患者外周血血MRP1表達(dá)與化療療效的研究[D];新疆醫(yī)科大學(xué);2004年

7 王雷;200例晚期非小細(xì)胞肺癌化療療效及預(yù)后影響因素的分析[D];大連醫(yī)科大學(xué);2013年

8 楊鳳嬌;MSCT增強(qiáng)掃描在進(jìn)展期胃癌早期化療療效評(píng)價(jià)中的價(jià)值[D];山西醫(yī)科大學(xué);2012年

9 吳狄;T細(xì)胞淋巴瘤組織微環(huán)境和COX-2表達(dá)與化療療效的相關(guān)性研究[D];安徽醫(yī)科大學(xué);2011年

10 楊寧;RRM1、ERCC1表達(dá)與晚期NSCLC患者GP方案化療療效及預(yù)后相關(guān)性分析[D];山東大學(xué);2011年

本文編號(hào)：2021143