TXR1通過(guò)調(diào)控TSP1、CD47及DUSP1參與鼻咽癌紫杉醇耐藥的研究
發(fā)布時(shí)間：2018-12-17 12:12
【摘要】：研究目的：紫杉醇是一種光譜抗腫瘤藥,廣泛應(yīng)用于多種惡性腫瘤,但是近年來(lái)紫杉醇耐藥現(xiàn)象的出現(xiàn)使化療效果大打折扣,紫杉醇耐藥機(jī)制的研究成為研究熱點(diǎn),但是諸多機(jī)制均不能系統(tǒng)闡明其耐藥機(jī)制。近年來(lái)發(fā)現(xiàn)了TXR1(紫杉醇耐藥基因1)為腫瘤研究提供新思路,但其功能及相關(guān)細(xì)胞內(nèi)信號(hào)通路仍不明確,本研究旨在前期研究基礎(chǔ)上進(jìn)一步探究TXR1基因的表達(dá)在鼻咽癌腫瘤細(xì)胞紫杉醇耐藥的功能。 研究方法：1.以人鼻咽癌紫杉醇耐藥細(xì)胞株CNE-l/Taxol為研究對(duì)象,用本課題組先前篩選的TXR1-22轉(zhuǎn)染該株細(xì)胞系,用RT-PCR及Western Blot方法確定TXR1基因是否被沉默； 2.采用MTT試驗(yàn)檢測(cè)耐藥細(xì)胞TXR1基因被沉默后,腫瘤耐藥能否逆轉(zhuǎn)； 3.TXR1-22轉(zhuǎn)染永生化正常鼻咽粘膜細(xì)胞NP69及鼻咽癌親本細(xì)胞CN E-1/親,采用MTT法檢測(cè)NP69及CNE-1/親生長(zhǎng)是否受抑制。 4.通過(guò)生物信息學(xué)技術(shù)篩選TSP1、 CD47、 DUSP1三種基因,在轉(zhuǎn)錄水平驗(yàn)證其與TXR1的關(guān)系 研究結(jié)果：1.RT-PCR及Western Blot結(jié)果顯示耐藥細(xì)胞株CNE-l/Taxol、中TXR1的表達(dá)明顯受抑制； 2.通過(guò)MTT法檢測(cè)了TXR1基因受抑制后腫瘤耐藥被部分逆轉(zhuǎn),且有統(tǒng)計(jì)學(xué)意義； 3.MTT法發(fā)現(xiàn)轉(zhuǎn)染前后的NP69及CNE-1/親生長(zhǎng)未受明顯抑制。 4.RT-PCR結(jié)果顯示TXR1基因沉默后TSP1、 CD47、 DUSP1三種基因轉(zhuǎn)錄水平表達(dá)明顯上調(diào),具有統(tǒng)計(jì)學(xué)意義 研究結(jié)論：1.通過(guò)RNA干擾技術(shù)同樣可以實(shí)現(xiàn)頭頸腫瘤相關(guān)基因的沉默； 2.在鼻咽癌耐藥細(xì)胞系的紫杉醇耐藥基因TXR1被沉默后,其耐藥性明顯降低； 3.TXR1基因可以作為逆轉(zhuǎn)紫杉醇耐藥的可能性靶點(diǎn),并且其對(duì)非靶向細(xì)胞無(wú)明顯毒性,為之后臨床研制新藥及安全性研究提供了體外實(shí)驗(yàn)依據(jù)； 4.TSP1、 CD47、 DUSP1為T(mén)XR1誘發(fā)紫杉醇耐藥的可能信號(hào)因子
[Abstract]:Objective: paclitaxel is a kind of spectral antitumor drug, which is widely used in many malignant tumors. However, the appearance of paclitaxel resistance in recent years has greatly reduced the effect of chemotherapy, and the mechanism of paclitaxel resistance has become a hot research topic. However, many mechanisms can not systematically elucidate the mechanism of drug resistance. In recent years, TXR1 (paclitaxel resistance gene 1) has been found to provide a new idea for tumor research, but its function and related intracellular signaling pathway are still unclear. The purpose of this study was to further explore the role of TXR1 gene expression in paclitaxel resistance in nasopharyngeal carcinoma (NPC) tumor cells. Methods: 1. The taxol resistant human nasopharyngeal carcinoma (NPC) cell line CNE-l/Taxol was transfected with previously screened TXR1-22. RT-PCR and Western Blot methods were used to determine whether the TXR1 gene was silenced. 2. MTT test was used to detect whether the drug resistance of tumor cells could be reversed after the TXR1 gene was silenced. 3.TXR1-22 was transfected into immortalized normal nasopharyngeal mucosal cells (NP69) and nasopharyngeal carcinoma parent cells (CN E-1 / parent). The growth inhibition of NP69 and CNE-1/ was detected by MTT assay. 4. Three genes of TSP1, CD47, DUSP1 were screened by bioinformatics, and the relationship between them and TXR1 was verified at the transcriptional level. The results of 1.RT-PCR and Western Blot showed that the expression of TXR1 in CNE-l/Taxol, was significantly inhibited. 2. MTT assay was used to detect the partial reversal of tumor resistance after TXR1 gene was inhibited, and 3.MTT method showed that the NP69 and CNE-1/ parental growth were not significantly inhibited before and after transfection. 4.RT-PCR results showed that the transcription level of three TSP1, CD47, DUSP1 genes was significantly up-regulated after TXR1 gene silencing, which was statistically significant: 1. The silencing of head and neck tumor-related genes can also be achieved by RNA interference technique. 2. The drug resistance of paclitaxel resistant gene TXR1 in nasopharyngeal carcinoma cell line was significantly decreased after the silencing of paclitaxel resistance gene. 3.TXR1 gene can be used as a possible target to reverse paclitaxel resistance, and it has no obvious toxicity to non-target cells, which provides in vitro experimental basis for clinical research of new drugs and safety. 4. TSP1, CD47, DUSP1 is the possible signal factor of paclitaxel resistance induced by TXR1.
【學(xué)位授予單位】：中南大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2014
【分類號(hào)】：R739.63
[Abstract]:Objective: paclitaxel is a kind of spectral antitumor drug, which is widely used in many malignant tumors. However, the appearance of paclitaxel resistance in recent years has greatly reduced the effect of chemotherapy, and the mechanism of paclitaxel resistance has become a hot research topic. However, many mechanisms can not systematically elucidate the mechanism of drug resistance. In recent years, TXR1 (paclitaxel resistance gene 1) has been found to provide a new idea for tumor research, but its function and related intracellular signaling pathway are still unclear. The purpose of this study was to further explore the role of TXR1 gene expression in paclitaxel resistance in nasopharyngeal carcinoma (NPC) tumor cells. Methods: 1. The taxol resistant human nasopharyngeal carcinoma (NPC) cell line CNE-l/Taxol was transfected with previously screened TXR1-22. RT-PCR and Western Blot methods were used to determine whether the TXR1 gene was silenced. 2. MTT test was used to detect whether the drug resistance of tumor cells could be reversed after the TXR1 gene was silenced. 3.TXR1-22 was transfected into immortalized normal nasopharyngeal mucosal cells (NP69) and nasopharyngeal carcinoma parent cells (CN E-1 / parent). The growth inhibition of NP69 and CNE-1/ was detected by MTT assay. 4. Three genes of TSP1, CD47, DUSP1 were screened by bioinformatics, and the relationship between them and TXR1 was verified at the transcriptional level. The results of 1.RT-PCR and Western Blot showed that the expression of TXR1 in CNE-l/Taxol, was significantly inhibited. 2. MTT assay was used to detect the partial reversal of tumor resistance after TXR1 gene was inhibited, and 3.MTT method showed that the NP69 and CNE-1/ parental growth were not significantly inhibited before and after transfection. 4.RT-PCR results showed that the transcription level of three TSP1, CD47, DUSP1 genes was significantly up-regulated after TXR1 gene silencing, which was statistically significant: 1. The silencing of head and neck tumor-related genes can also be achieved by RNA interference technique. 2. The drug resistance of paclitaxel resistant gene TXR1 in nasopharyngeal carcinoma cell line was significantly decreased after the silencing of paclitaxel resistance gene. 3.TXR1 gene can be used as a possible target to reverse paclitaxel resistance, and it has no obvious toxicity to non-target cells, which provides in vitro experimental basis for clinical research of new drugs and safety. 4. TSP1, CD47, DUSP1 is the possible signal factor of paclitaxel resistance induced by TXR1.
【學(xué)位授予單位】：中南大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2014
【分類號(hào)】：R739.63
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