發(fā)布時(shí)間：2018-08-02 09:07

【摘要】：研究背景：肺癌是目前世界上最常見的惡性腫瘤,死亡率位居惡性腫瘤之首。隨著基因組學(xué)的發(fā)展及靶向藥物的開發(fā)和應(yīng)用,晚期非小細(xì)胞肺癌(non-small cell lung cancer,NSCLC)的一線治療正由傳統(tǒng)的化療轉(zhuǎn)變?yōu)閭�(gè)體化的分子靶向治療。表皮生長(zhǎng)因子受體(epidermal growth factor receptor,EGFR)酪氨酸激酶抑制劑(tyrosine kinase inhibitors,TKI)在目前的NSCLC的治療中得到了廣泛應(yīng)用,尤其不吸煙的亞洲女性患者及存在EGFR突變的患者對(duì)EGFR-TKI治療效果尤為顯著。多項(xiàng)臨床研究包括EURTAC.WJTOG340、 IPASS、NEJ0025等，，已證實(shí)EGFR-TKI作為一線藥物用于治療EGFR存在敏感突變的晚期NSCLC患者,其效果優(yōu)于化療。然而,一些NSCLC患者對(duì)EGFR-TKI存在原發(fā)性耐藥現(xiàn)象。另外,即便是對(duì)EGFR-TKI治療有效的患者,絕大多數(shù)也會(huì)在一年內(nèi)出現(xiàn)獲得性耐藥。所以,對(duì)于接受分子靶向治療的肺癌患者,無(wú)論對(duì)EGFR-TKI治療是否有效,最終都難以逃脫面疾病進(jìn)展及復(fù)發(fā)的厄運(yùn)。EGFR-TKI的耐藥問(wèn)題目前已經(jīng)成為NSCLC靶向治療臨床應(yīng)用的主要瓶頸,只有明確了EGFR-TKI的耐藥原因和機(jī)制，才能夠找到克服耐藥的方法,從而使更多的NSCLC患者從分子靶向治療中獲益。根據(jù)目前研究,T790M突變以及MET擴(kuò)增是最為常見的兩種獲得性耐藥機(jī)制,二者約占到所有獲得性耐藥的60%。另外40%的機(jī)制尚未完全闡明,其中研究相對(duì)較多的為張力蛋白同源10號(hào)染色體缺失的磷酸酶(Phosphatase and tensin homolog deleted on chromosome ten,PTEN)抑癌基因的缺失。PTEN是目前為止發(fā)現(xiàn)的第一個(gè)具有磷酸酶活性的抑癌基因,其表達(dá)產(chǎn)物在PI3K/Akt信號(hào)通路中發(fā)揮著重要的作用。它可以抑制下游信號(hào)通路中Akt的活化,控制細(xì)胞的增殖,從而起到抑制腫瘤生長(zhǎng)的作用。研究表明PTEN在多種惡性腫瘤細(xì)胞系中(包括肺癌、食管癌、肝癌等)處于低表達(dá)狀態(tài),并對(duì)腫瘤的發(fā)生發(fā)展起重要作用。國(guó)外Kokubo等人的研究表明PTEN低表達(dá)與吉非替尼獲得性耐藥相關(guān)。Endoh等人的研究結(jié)果也表明了在接受吉非替尼治療的患者中PTEN表達(dá)水平較高的患者預(yù)后較好,而EGFR突變的患者若存在PTEN缺失,EGFR-TKI治療效果不佳。Sos等人的研究顯示,PTEN可通過(guò)激活A(yù)kt導(dǎo)致EGFR-TKI耐藥。Zhuang等人在2009年通過(guò)X線照射的方式上調(diào)了H-157肺癌細(xì)胞PTEN的表達(dá)水平,增加了癌細(xì)胞對(duì)吉非替尼的敏感性。以上研究均表明PTEN在T790M突變以及MET擴(kuò)增以外的EGFR-TKI耐藥機(jī)制中扮演了相當(dāng)重要的角色。2006年Li等人在研究針對(duì)DNA啟動(dòng)子序列的dsRNA對(duì)基因表達(dá)調(diào)控的過(guò)程中發(fā)現(xiàn),針對(duì)啟動(dòng)子區(qū)的非CpG島序列為靶點(diǎn)的dsRNA可以引起某些表達(dá)水平較低的基因的表達(dá)增高,并將這種現(xiàn)象定義RNA激活(RNAActivation,RNAa)。他將具有RNA激活功能的小RNA分子稱為“小激活RNA”(small activating RNA,saRNA)。RNAa與RNA干攏(RNA interfering,RNAi)對(duì)目標(biāo)基因表達(dá)的影響,一個(gè)是增強(qiáng),一個(gè)是抑制,二者均是通過(guò)向細(xì)胞內(nèi)導(dǎo)人小片段雙鏈RNA而發(fā)揮作用,都有高效、特異、相對(duì)容易操作的特點(diǎn)。RNAa現(xiàn)象的發(fā)現(xiàn)為腫瘤的基因治療提供了新的策略。RNAa能夠特異性地激活細(xì)胞內(nèi)已經(jīng)存在的表達(dá)水平較低的基因,在應(yīng)用于治療腫瘤時(shí),不需要考慮腫瘤是否存在特定的致癌基因,也不需要導(dǎo)人新的基因。因此我們能夠更加容易的使某個(gè)低表達(dá)的基因特異性增強(qiáng),具有獨(dú)特的優(yōu)勢(shì)。我們可以利用RNAa激活腫瘤抑制基因、細(xì)胞凋亡基因以及細(xì)胞周期抑制基因等來(lái)治療腫瘤。Li等人分析了p21Waf1/Cip1(p21)和人鈣粘連素E基因啟動(dòng)子結(jié)構(gòu),在該序列的非CpG島序列上篩選出能激活下游基因轉(zhuǎn)錄的saRNA作用于靶位點(diǎn)。他合成了長(zhǎng)度為21個(gè)核苷酸的saRNA,然后將該saRNA轉(zhuǎn)染到前列腺癌、宮頸癌、人膀胱癌、和乳腺癌等多種體外腫瘤細(xì)胞中,結(jié)果發(fā)現(xiàn)轉(zhuǎn)染了saRNA分子的腫瘤細(xì)胞與對(duì)照組細(xì)胞相比,鈣粘連素E和p21基因的表達(dá)水平在不同的細(xì)胞出現(xiàn)了2～13倍的增加,腫瘤細(xì)胞的生長(zhǎng)受到顯著抑制。這些研究雖然多數(shù)是體外實(shí)驗(yàn),但已經(jīng)取得了較好的成果,預(yù)示RNAa技術(shù)在腫瘤基因治療上具有極為良好的發(fā)展前景。PTEN在癌細(xì)胞中的缺失、低表達(dá)與EGFR-TKI的耐藥密切相關(guān),目前尚未有利用RNA激活技術(shù)上調(diào)肺癌PTEN表達(dá)的相關(guān)研究。鑒于此,本研究選擇PTEN為靶點(diǎn)，設(shè)計(jì)合成針對(duì)PTEN基因啟動(dòng)子的saRNA來(lái)進(jìn)實(shí)驗(yàn),旨在探討RNAa上調(diào)PTEN基因表達(dá)克服EGFR-TKI耐藥的可行性及有效性為EGFR-TKI耐藥后的治療尋找新的方法及思路。第一部分PTEN表達(dá)與非小細(xì)胞肺癌TKI耐藥關(guān)系的研究目的：通過(guò)對(duì)人非小細(xì)胞肺癌H-157、H-1355、H-1650三種細(xì)胞PTEN、 pAkt的表達(dá)強(qiáng)度及三種細(xì)胞對(duì)TKI的療效進(jìn)行分析，探討PTEN低表達(dá)或表達(dá)缺失對(duì)TKI耐藥的影響,并研究其導(dǎo)致耐藥的可能機(jī)制。方法：使用逆轉(zhuǎn)錄PCR及Western blot對(duì)H-157、H-1355、H-1650三種肺癌細(xì)胞進(jìn)行檢測(cè)分別從RNA和蛋白水平評(píng)估PTEN的表達(dá)強(qiáng)度。繪制TKI處理前后的三種細(xì)胞的生長(zhǎng)曲線并使用流式細(xì)胞儀檢測(cè)細(xì)胞凋亡的情況，評(píng)價(jià)TKI對(duì)三種細(xì)胞的療效。使用Western blot檢測(cè)TKI處理后的三種肺癌細(xì)胞pAkt的表達(dá)情況以探討PTEN低表達(dá)導(dǎo)致TKI耐藥的機(jī)制。結(jié)果：H-1355細(xì)胞PTEN表達(dá)強(qiáng)度最高,H-157細(xì)胞表達(dá)強(qiáng)度較低,而H-1650細(xì)胞中PTEN蛋白表達(dá)缺失經(jīng)過(guò)TKI藥物處理后H-1355的生長(zhǎng)曲線明顯下移,細(xì)胞有明顯的凋亡(p0.05),H-1650細(xì)胞的生長(zhǎng)曲線略有下移,未見有明顯的細(xì)胞凋亡(p0.05),H-157細(xì)胞的生長(zhǎng)曲線沒(méi)有明顯變化,未見有明顯的細(xì)胞凋亡(p0.05)：經(jīng)過(guò)TKI處理后H-1355細(xì)胞pAkt呈現(xiàn)低表達(dá)，H-157及H-1650都呈現(xiàn)高表達(dá)狀態(tài)。結(jié)論：TKI的耐藥與PTEN的表達(dá)強(qiáng)度有一定的關(guān)系。PTEN表達(dá)水平正常的肺癌細(xì)胞對(duì)TKI治療較為敏感,而PTEN缺失或者PTEN表達(dá)明顯下降的肺癌細(xì)胞對(duì)TKI治療效果不明顯。因此PTEN的低表達(dá)或者表達(dá)缺失可能是導(dǎo)致肺癌對(duì)TKI耐藥的機(jī)制之一。而PTEN低表達(dá)或表達(dá)缺失導(dǎo)致TKI耐藥的原因可能與其所調(diào)控的Akt/PI3K通路的持續(xù)活化有關(guān)。第二部分針對(duì)PTEN基因的小激活RNA的構(gòu)建與篩選目的：針對(duì)PTEN基因,設(shè)計(jì)并篩選出具有明顯激活功能的saRNAo方法：根據(jù)現(xiàn)有文獻(xiàn)報(bào)道的saRNA設(shè)計(jì)原則,設(shè)計(jì)5條dsRNA作為候選saRNA,交由上海生工公司化學(xué)合成后轉(zhuǎn)染人人非小細(xì)胞肺癌H-157細(xì)胞。通過(guò)RT-PCR檢測(cè)轉(zhuǎn)染后細(xì)胞內(nèi)PTEN的表達(dá)強(qiáng)度,篩選出具有激活功能的saRNAo然后將篩選出的功能性saRNA轉(zhuǎn)染至H-157細(xì)胞,使用Western blot檢測(cè)PTEN蛋白水平的表達(dá)強(qiáng)度,評(píng)估saRNA的激活作用是否適用于肺組織細(xì)胞中。結(jié)果：我們?cè)O(shè)計(jì)出的5條候選dsRNA中有3條可以使H-157細(xì)胞中的PTEN表達(dá)上調(diào),其中的一條可以使PTEN的表達(dá)強(qiáng)度增強(qiáng)2倍以上。將該saRNA轉(zhuǎn)染人H-157細(xì)胞后,可以從RNA及蛋白水平增強(qiáng)PTEN的表達(dá)。結(jié)論：RNA激活現(xiàn)象同樣存在于肺組織細(xì)胞中,通過(guò)向肺癌細(xì)胞內(nèi)轉(zhuǎn)人針對(duì)PTEN的功能性saRNA,可以特異性的將PTEN的表達(dá)上調(diào)。第三部分saRNA上調(diào)PTEN表達(dá)對(duì)肺癌TKI耐藥的影響目的：通過(guò)將篩選出的saRNA轉(zhuǎn)染人H-157細(xì)胞上調(diào)PTEN的表達(dá),觀察PTEN的表達(dá)上調(diào)能否逆轉(zhuǎn)TKI的耐藥狀態(tài)。方法：將篩選出的功能性saRNA轉(zhuǎn)染至H-157細(xì)胞,使用RT-PCR及Western blot方法分別從RNA及蛋白水平檢測(cè)PTEN的表達(dá)強(qiáng)度，評(píng)估saRNA能否使PTEN的表達(dá)上調(diào)。同時(shí)通過(guò)細(xì)胞生長(zhǎng)曲線及細(xì)胞凋亡的檢測(cè)觀察轉(zhuǎn)染saRNA后的H-157細(xì)胞對(duì)TKI藥物的敏感性,評(píng)估PTEN的表達(dá)上調(diào)能否克服TKI耐藥。結(jié)果：saRNA轉(zhuǎn)染至H-157細(xì)胞后,RT-PCR及Western blot檢測(cè)結(jié)果均顯示PTEN的表達(dá)水平較dsControl組及mock組明顯增強(qiáng)(p0.05),這說(shuō)明saRNA在H-157細(xì)胞內(nèi)發(fā)揮了RNA激活作用使得PTEN的表達(dá)上調(diào)。生長(zhǎng)曲線顯示轉(zhuǎn)染saRNA 72小時(shí)后,saRNA組細(xì)胞的生長(zhǎng)明顯受抑制,與其他兩組差異有顯著統(tǒng)計(jì)學(xué)意義(p0.05)。細(xì)胞凋亡結(jié)果顯示saRNA組細(xì)胞凋亡率為17.82±2.37%,dsControl組細(xì)胞凋亡率為1.74+0.17%,mock組細(xì)胞凋亡率為0.33+0.14%(p0.05)。結(jié)論:PTEN的低表達(dá)是造成TKI耐藥的機(jī)制之一,導(dǎo)致耐藥的原因是PTEN的低表達(dá)造成Akt的過(guò)度磷酸化,從而使Akt/PI3K通路活化,引起耐藥。通過(guò)篩選出的功能性saRNA可以上調(diào)PTEN的表達(dá)水平,從而增加肺癌細(xì)胞對(duì)TKI的敏感性,逆轉(zhuǎn)TKI的耐藥狀態(tài)。
[Abstract]:Background: lung cancer is the most common malignant tumor in the world and the death rate is the first in malignant tumor. With the development of genomics and the development and application of targeted drugs, the first-line treatment of advanced non small cell lung cancer (non-small cell lung cancer, NSCLC) is changing from traditional chemotherapy to individualized molecular targeting therapy. The growth factor receptor (epidermal growth factor receptor, EGFR) tyrosine kinase inhibitor (tyrosine kinase inhibitors, TKI) has been widely used in the current treatment of NSCLC, especially in non smoking Asian women and those with EGFR mutations, which are particularly effective for EGFR-TKI treatment. G340, IPASS, NEJ0025, and so on, have proved EGFR-TKI as a first-line drug for the treatment of EGFR with sensitive mutations in late NSCLC patients, the effect is better than chemotherapy. However, some NSCLC patients have primary drug resistance to EGFR-TKI. In addition, the overwhelming majority of patients who are effective for EGFR-TKI treatment will also have acquired resistance within one year. Therefore, for patients with lung cancer receiving molecular targeted therapy, whether the treatment of EGFR-TKI is effective or not, it is difficult to escape the problem of resistance to the progression of the face disease and the relapse of the drug-resistant.EGFR-TKI, which has become the main bottleneck in the clinical application of the target therapy of NSCLC. Only the reason and mechanism of the drug resistance of EGFR-TKI can be found to be overcome. The drug resistance method makes more NSCLC patients benefit from molecular targeting therapy. According to current research, T790M mutation and MET amplification are the two most common mechanisms of acquired resistance. The two other 40% mechanisms for all acquired resistance to 60%. have not been fully elucidated, and more of them are tenmifying protein homologous 10. The deletion of the phosphatase (Phosphatase and tensin homolog deleted on chromosome ten, PTEN) the deletion of the tumor suppressor gene is the first tumor suppressor gene found with the activity of phosphatase so far. Its expression product plays an important role in the PI3K/Akt signaling pathway. It can inhibit the downstream signal pathway in the Akt. The study shows that PTEN is in a low expression state in a variety of malignant tumor cell lines (including lung, esophageal and liver cancer), and plays an important role in the development of tumor. The study of foreign Kokubo et al. Shows that the low expression of PTEN is associated with the acquired resistance of gefitinib. .Endoh et al's results also suggest that patients with high PTEN expression in gefitinib patients have a better prognosis, while EGFR mutations in patients with PTEN deletion and EGFR-TKI treatment are not effective in.Sos et al. Studies show that PTEN can lead to EGFR-TKI - resistant.Zhuang and others through X - ray exposure in 2009 by activating Akt The expression level of PTEN in H-157 lung cancer cells increased and the sensitivity of cancer cells to gefitinib was increased. All of these studies showed that PTEN played a significant role in the mechanism of T790M mutation and EGFR-TKI resistance other than MET amplification,.2006 Li et al. In the study of the gene expression regulation of dsRNA for the DNA promoter sequence. It was found that dsRNA, the target of the non CpG Island sequence of the promoter region, could cause a higher expression of some genes with lower levels of expression, and defined the phenomenon as RNA activation (RNAActivation, RNAa). He called the small RNA molecule with the RNA activation function as "small activated RNA" (small activating RNA, saRNA) and dried together. The effects of interfering, RNAi) on the expression of target genes, one is enhancement, one is inhibition, and the two are both effective, specific, and relatively easy to operate by the small fragment of double stranded RNA. The discovery of the.RNAa phenomenon provides a new strategy for the treatment of tumor gene therapy,.RNAa can specifically activate the cells. In the treatment of tumors, a gene with a lower level of expression is not required to consider whether a tumor has a specific oncogene or a new gene. Therefore, we can make a particular gene specific enhancement of a low expression more unique. We can use RNAa to activate tumor inhibition. Gene, apoptosis gene, and cell cycle inhibition gene were used to treat tumor.Li and others to analyze the promoter structure of p21Waf1/Cip1 (p21) and human cadherin E gene, and to screen out saRNA that can activate downstream gene transcription on the target site on the non CpG Island sequence of this sequence. He synthesized the saRNA of 21 nucleotides, and then The saRNA was transfected into a variety of extracorporeal tumor cells, such as prostate cancer, cervical cancer, human bladder cancer, and breast cancer. The results showed that the expression level of saRNA and the expression level of calcin E and p21 gene increased 2~13 times in different cells compared with the control group, and the growth of tumor cells was significantly inhibited. Although most of the studies are in vitro experiments, good results have been achieved, which indicates that RNAa technology has a very good development prospect in tumor gene therapy, the lack of.PTEN in cancer cells, low expression is closely related to the drug resistance of EGFR-TKI. At present, there is no related research on the use of RNA activation technology to increase the expression of PTEN in lung cancer. In this study, we selected PTEN as the target to design and synthesize the saRNA for the PTEN gene promoter, aiming at exploring the feasibility and effectiveness of RNAa up regulation of PTEN gene expression to overcome EGFR-TKI resistance and finding new ways and ideas for the treatment of EGFR-TKI after drug resistance. The first part of the study of the relationship between PTEN expression and TKI resistance in non-small cell lung cancer By analyzing the expression intensity of three cells of human non-small cell lung cancer (H-157, H-1355, H-1650), the expression intensity of pAkt and the effect of three kinds of cells on TKI, the effect of PTEN low expression or expression loss on the drug resistance of TKI was investigated and the possible mechanism of drug resistance was studied. Method: to make use of reverse transcriptase PCR and Western blot to H-157, three The expression intensity of PTEN was evaluated from RNA and protein levels. The growth curve of three cells before and after TKI treatment was plotted and the apoptosis was detected by flow cytometry. The effect of TKI on three kinds of cells was evaluated. The expression of pAkt in three lung cancer cells treated with TKI was detected by Western blot The mechanism of low expression of PTEN led to TKI resistance. Results: the expression of PTEN in H-1355 cells was the highest, and the expression intensity of H-157 cells was low, while the growth curve of PTEN protein expression in H-1650 cells was obviously down after TKI drug treatment, and the cells had obvious apoptosis (P0.05), and the growth curve of H-1650 cells was slightly down, and there was no obvious change in the growth curve of H-1650 cells. There was no obvious change in the growth curve of H-157 cells and no obvious cell apoptosis (P0.05): pAkt showed low expression in H-1355 cells after TKI treatment, and both H-157 and H-1650 showed high expression state. Conclusion: the resistance of TKI to PTEN expression intensity has certain relation to the normal lung cancer cells with.PTEN expression level. It is more sensitive to TKI therapy, but the effect of PTEN deletion or the decreased expression of PTEN on TKI is not obvious. Therefore, the low expression of PTEN or the lack of expression may be one of the mechanisms that lead to the resistance of lung cancer to TKI. The reason that the low expression of PTEN or the deletion of the expression causes the drug resistance of TKI may be related to the holding of the Akt/PI3K pathway controlled by the PTEN. The second part is aimed at the construction and screening of the small activation RNA of PTEN gene: designed and screened the saRNAo method with obvious activation function for the PTEN gene: according to the saRNA design principle reported in the existing literature, the 5 dsRNA is designed as the candidate saRNA, and the transfection of the Shanghai raw material company after chemical synthesis is not small. Cell lung cancer H-157 cells. The expression intensity of PTEN in the cells after transfection was detected by RT-PCR. The activation function was screened and then the functional saRNA was transfected into H-157 cells. The expression intensity of PTEN protein level was detected by Western blot, and the activation of saRNA was evaluated in lung tissue cells. 3 of the 5 candidate dsRNA we designed can increase the expression of PTEN in H-157 cells, one of which can increase the expression intensity of PTEN by more than 2 times. After transfection of the saRNA to human H-157 cells, the expression of PTEN can be enhanced from RNA and protein levels. Conclusion: RNA activation is also present in lung tissue cells, through the lung. The tumor cells transferred to the functional saRNA of PTEN, the expression of PTEN can be specifically up-regulated. Third part saRNA up-regulated the effect of PTEN expression on the drug resistance of lung cancer. By using the selected saRNA transfected human H-157 cells to up-regulation the expression of PTEN, whether the up regulation of PTEN expression can reverse the TKI drug resistance state. The functional saRNA was transfected into H-157 cells, and the expression intensity of PTEN was detected from RNA and protein levels by RT-PCR and Western blot methods. The expression of PTEN was up regulated by saRNA, and the sensitivity of H-157 cell to TKI drugs was evaluated through the detection of cell growth curve and apoptosis. TKI resistance was overcome. Results: after saRNA transfection to H-157 cells, the results of RT-PCR and Western blot showed that the expression level of PTEN was significantly higher than that of dsControl and mock groups (P0.05), which indicated that saRNA in the H-157 cells was activated to make the expression up. The growth curve showed that the transfection was 72 hours after transfection. The growth of the group cells was obviously inhibited, and there was significant difference with the other two groups (P0.05). Apoptosis results showed that the apoptosis rate of group saRNA was 17.82 + 2.37%, the apoptosis rate of group dsControl was 1.74+0.17%, and the apoptosis rate of mock group was 0.33+0.14% (P0.05). Conclusion: the low expression of PTEN is one of the mechanisms causing TKI resistance. The reason for the resistance is that the low expression of PTEN causes excessive phosphorylation of Akt, which causes the activation of Akt/PI3K pathway and causes resistance. Through the screening of functional saRNA, the expression level of PTEN can be up-regulated, thus increasing the sensitivity of lung cancer cells to TKI and reversing the drug resistance of TKI.
【學(xué)位授予單位】：山東大學(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 魯慧英;Detection of hepatitis C virus RNA sequences in cholangiocarcinomas in Chinese and American patients[J];Chinese Medical Journal;2000年12期

2 ;Detection the HCV RNA by PCR-microplate hybridization method from clinical specimens[J];中國(guó)輸血雜志;2001年S1期

3 付漢江,鄭曉飛;類mRNA RNA研究進(jìn)展[J];國(guó)外醫(yī)學(xué)(分子生物學(xué)分冊(cè));2002年04期

4 Verheyden B ,徐其武;口服脊髓灰質(zhì)炎疫苗核殼和RNA的穩(wěn)定性[J];國(guó)外醫(yī)學(xué).預(yù)防.診斷.治療用生物制品分冊(cè);2002年01期

5 CMBE譯文組;探索小RNA的功能[J];現(xiàn)代臨床醫(yī)學(xué)生物工程學(xué)雜志;2004年03期

6 沈維干;RNA interference and its current application in mammals[J];Chinese Medical Journal;2004年07期

7 ;Potent and specific inhibition of SARS-CoV antigen expression by RNA interference[J];Chinese Medical Journal;2005年09期

8 孫娣;汪洋;張麗娟;閆玉清;;一種簡(jiǎn)捷提取植物總RNA的方法[J];黑龍江醫(yī)藥;2005年06期

9 熊慧華;于世英;胡廣原;莊亮;;Effects of Survivin Expression Suppressed by Short Hairpin RNA on MCF-7 Cells[J];華中科技大學(xué)學(xué)報(bào)(醫(yī)學(xué)英德文版);2006年03期

10 楊益超;;RNA干擾及其在醫(yī)學(xué)中的應(yīng)用[J];廣西醫(yī)學(xué);2007年10期

相關(guān)會(huì)議論文 前10條

1 金由辛;;面向21世紀(jì)的RNA研究[A];面向21世紀(jì)的科技進(jìn)步與社會(huì)經(jīng)濟(jì)發(fā)展（下冊(cè)）[C];1999年

2 ;第四屆RNA全國(guó)研討會(huì)大會(huì)報(bào)告日程安排[A];第四屆全國(guó)RNA進(jìn)展研討會(huì)論文集[C];2005年

3 ;Function of Transfer RNA Modifications in Plant Development[A];植物分子生物學(xué)與現(xiàn)代農(nóng)業(yè)——全國(guó)植物生物學(xué)研討會(huì)論文摘要集[C];2010年

4 王峰;張秋平;陳金湘;;棉花總RNA的快速提取方法[A];中國(guó)棉花學(xué)會(huì)2011年年會(huì)論文匯編[C];2011年

5 關(guān)力;陳本iY;iJ云虹;郭培芝;魏重琴;邱蘇吾;苗健;;關(guān)于動(dòng)物}D~T中RNAn,定方法的研究[A];中國(guó)生理科學(xué)會(huì)學(xué)術(shù)會(huì)議論文摘要匯編（生物化學(xué)）[C];1964年

6 夏海濱;;小RNA在免疫學(xué)領(lǐng)域中的應(yīng)用研究進(jìn)展[A];中國(guó)免疫學(xué)會(huì)第五屆全國(guó)代表大會(huì)暨學(xué)術(shù)會(huì)議論文摘要[C];2006年

7 ;The stability of hepatitis C virus RNA in various handling and storage conditions[A];中國(guó)輸血協(xié)會(huì)第四屆輸血大會(huì)論文集[C];2006年

8 郭德銀;;RNA干擾在病毒研究和控制中的應(yīng)用[A];2006中國(guó)微生物學(xué)會(huì)第九次全國(guó)會(huì)員代表大會(huì)暨學(xué)術(shù)年會(huì)論文摘要集[C];2006年

9 甘儀梅;楊業(yè)華;王學(xué)奎;曹燕;楊特武;;棉花總RNA快速提取[A];中國(guó)棉花學(xué)會(huì)2007年年會(huì)論文匯編[C];2007年

10 ;Identification and characterization of novel interactive partner proteins for PCBP1 that is a RNA-binding protein[A];中國(guó)優(yōu)生優(yōu)育協(xié)會(huì)第四屆全國(guó)學(xué)術(shù)論文報(bào)告會(huì)暨基因科學(xué)高峰論壇論文專輯[C];2008年

相關(guān)重要報(bào)紙文章 前10條

1 記者 馮衛(wèi)東;研究人員發(fā)現(xiàn)可破壞腫瘤抑制基因的小RNA[N];科技日?qǐng)?bào);2009年

2 記者 儲(chǔ)笑抒 通訊員 盛偉;人體微小RNA有望提前發(fā)出癌癥預(yù)警[N];南京日?qǐng)?bào);2011年

3 瀘州醫(yī)學(xué)院副教授、科普作家 周志遠(yuǎn);“大頭兒子”與環(huán)狀RNA[N];第一財(cái)經(jīng)日?qǐng)?bào);2014年

4 麥迪信;小分子RNA可能有大作用[N];醫(yī)藥經(jīng)濟(jì)報(bào);2003年

5 董映璧;美發(fā)現(xiàn)基因調(diào)控可回應(yīng)“RNA世界”[N];科技日?qǐng)?bào);2006年

6 張忠霞;特制RNA輕推一下,就能“喚醒”基因[N];新華每日電訊;2007年

7 聶翠蓉;RNA：縱是配角也精彩[N];科技日?qǐng)?bào);2009年

8 馮衛(wèi)東;RNA干擾機(jī)制首次在人體中獲得證實(shí)[N];科技日?qǐng)?bào);2010年

9 馮衛(wèi)東　王小龍;英在地球早期環(huán)境模擬條件下合成類RNA[N];科技日?qǐng)?bào);2009年

10 記者 常麗君;新技術(shù)讓研究進(jìn)入單細(xì)胞內(nèi)RNA的世界[N];科技日?qǐng)?bào);2011年

相關(guān)博士學(xué)位論文 前10條

1 王趙瑋;昆蟲RNA病毒復(fù)制及昆蟲抗病毒天然免疫機(jī)制研究[D];武漢大學(xué);2014年

2 包純;一類新非編碼RNA的發(fā)現(xiàn)以及產(chǎn)生和功能的初探[D];華中師范大學(xué);2015年

3 李語(yǔ)麗;基于MeRIP-seq的水稻RNA m6A甲基化修飾的研究[D];中國(guó)科學(xué)院北京基因組研究所;2015年

4 熊瑜琳;miR-122靶位基因STAT3調(diào)控長(zhǎng)鏈非編碼 RNA Lethe促進(jìn)HCV復(fù)制的機(jī)制研究[D];第三軍醫(yī)大學(xué);2015年

5 范春節(jié);高通量測(cè)序鑒定毛竹小RNA及其功能分析[D];中國(guó)林業(yè)科學(xué)研究院;2012年

6 王加強(qiáng);小鼠著床前胚胎特異ERV相關(guān)長(zhǎng)非編碼RNA的定向篩選及功能研究[D];東北農(nóng)業(yè)大學(xué);2015年

7 王業(yè)偉;非編碼RNA SPIU的結(jié)構(gòu)和功能研究和p19INK4D在APL發(fā)病中的作用[D];上海交通大學(xué);2013年

8 鄒艷芬;子癇前期中非編碼RNA對(duì)滋養(yǎng)細(xì)胞功能的調(diào)控及機(jī)制探索[D];南京醫(yī)科大學(xué);2015年

9 朱喬;miR-10b在人肝細(xì)胞肝癌發(fā)生中的作用及其機(jī)制的初步探索[D];第四軍醫(yī)大學(xué);2015年

10 蔣俊鋒;長(zhǎng)鏈非編碼RNA BACE1-AS促進(jìn)Aβ聚集及其調(diào)節(jié)BACE1和SERF1a的ceRNA機(jī)制研究[D];第二軍醫(yī)大學(xué);2015年

相關(guān)碩士學(xué)位論文 前10條

1 陸聰兒;條紋斑竹鯊再生肝臟中差異RNA的研究[D];浙江理工大學(xué);2015年

2 張曉輝;小鼠幾種長(zhǎng)鏈非編碼RNA基因功能的初步研究[D];昆明理工大學(xué);2015年

3 全弘揚(yáng);長(zhǎng)鏈非編碼RNA在細(xì)胞內(nèi)質(zhì)網(wǎng)應(yīng)激反應(yīng)中的相關(guān)作用及機(jī)制研究[D];北京協(xié)和醫(yī)學(xué)院;2015年

4 胡亮;DDX19A識(shí)別PRRSV基因組RNA并激活NLRP3炎癥小體[D];中國(guó)農(nóng)業(yè)科學(xué)院;2015年

5 張帥兵;應(yīng)用RNA干擾技術(shù)對(duì)旋毛蟲Nudix水解酶基因功能的研究[D];鄭州大學(xué);2015年

6 鄭芳芳;肺癌RNA-Seq數(shù)據(jù)中RNA編輯事件的識(shí)別算法和系統(tǒng)分析[D];蘇州大學(xué);2015年

7 陳瑞東;長(zhǎng)鏈非編碼RNA MEG3在胰腺癌發(fā)生發(fā)展中作用的實(shí)驗(yàn)研究[D];蘇州大學(xué);2015年

8 劉駿武;線蟲和水稻中環(huán)狀RNA的預(yù)測(cè)及分析[D];華中農(nóng)業(yè)大學(xué);2015年

9 李猷;利用RNA干擾技術(shù)提高番茄抗TMV侵染能力的研究[D];牡丹江師范學(xué)院;2015年

10 吳術(shù)盛;SVCV感染EPC細(xì)胞microRNA表達(dá)譜的初步研究[D];華中農(nóng)業(yè)大學(xué);2015年

本文編號(hào)：2158944