【摘要】：癌變是一個復(fù)雜的過程,是由癌基因激活,抑癌基因和細(xì)胞死亡機制失活相互作用而出現(xiàn)的。由于細(xì)胞凋亡的減少與腫瘤發(fā)生密切相關(guān),這提示凋亡過程中起負(fù)調(diào)控有關(guān)的基因可能有致癌性,而促進(jìn)細(xì)胞凋亡有關(guān)的基因可能為腫瘤抑制基因。因此,凋亡相關(guān)蛋白一直是腫瘤研究的熱點領(lǐng)域。本文將分章討論一新穎抗凋亡蛋白ARC與鼻咽癌發(fā)生、發(fā)展的關(guān)系及對鼻咽癌放療與化療敏感性的影響。 目的蛋白水平探討鼻咽癌組織及鼻咽部炎癥組織中ARC的表達(dá)與腫瘤T分期、臨床分期和有無淋巴結(jié)轉(zhuǎn)移等臨床病理學(xué)參數(shù)的關(guān)系,并檢測ARC在鼻咽癌細(xì)胞系中的表達(dá)水平及細(xì)胞內(nèi)定位。 方法采用免疫組織化學(xué)的方法檢測82例鼻咽癌組織、20例鼻咽部慢性炎癥組織的石蠟組織標(biāo)本,分析其在鼻咽癌組織中的表達(dá)與臨床病理學(xué)參數(shù)的關(guān)系。采用細(xì)胞免疫組化檢測ARC在鼻咽癌細(xì)胞株中的表達(dá)定位,Western blot實驗方法檢測ARC在鼻咽癌細(xì)胞株中的表達(dá)。SPSS17.0統(tǒng)計軟件對實驗數(shù)據(jù)進(jìn)行統(tǒng)計分析。 結(jié)果 1)通過鼻咽癌組織及細(xì)胞的免疫組化檢測,結(jié)果發(fā)現(xiàn)ARC表達(dá)于細(xì)胞漿及細(xì)胞核內(nèi),其中大部分表達(dá)于細(xì)胞漿。 2)ARC蛋白在鼻咽癌組織中的表達(dá)強度明顯高于鼻咽部炎癥組織(P0.001)。其中在鼻咽癌組織中,44例為強陽性(53.7%)23例為中度陽性(28%),15例為弱陽性(18.3%)；鼻咽部慢性炎癥組織中,16例為弱陽性(80%),4例為中度陽性(20%),無強陽性。 3)ARC蛋白的表達(dá)與鼻咽癌的臨床分期、病理分型因素相關(guān)(P0.001),臨床分期越晚、分化程度越低的組織中ARC的表達(dá)越高。而與患者年齡、性別及有無淋巴結(jié)轉(zhuǎn)移無關(guān)(P0.05)。 4)ARC在鼻咽癌細(xì)胞株CNE-1、CNE-2、5-8F、6-10B中均有明顯表達(dá),而在鼻咽永生化上皮細(xì)胞NP69中表達(dá)最弱；其相對表達(dá)量分別為CNE-1(4.78±0.47)、CNE-2(5.39±0.58)、5-8F (3.54±0.33)、6-10B (3.12±0.26)、NP69(1.67±0.24)。在鼻咽癌細(xì)胞株中表達(dá)的表達(dá)量由高到低依次為CNE-2CNE-15-8F6-10B。 結(jié)論以上結(jié)果提示ARC在鼻咽癌的發(fā)生、發(fā)展中可能起重要作用,臨床上檢測ARC的表達(dá)對鼻咽癌惡性程度的預(yù)測可能會有所幫助。 目的使用RNA干擾技術(shù)及基因克隆技術(shù)分別沉默和過表達(dá)ARC在鼻咽癌細(xì)胞株中的表達(dá),并篩選出穩(wěn)定轉(zhuǎn)染的細(xì)胞株。 方法使用pcDNA6.2-GW/EmGFP/miRNA載體構(gòu)建表達(dá)miRNA對ARC基因的RNA干擾的質(zhì)粒；使用pEZ-M03載體構(gòu)建表達(dá)ARC基因的質(zhì)粒。采用脂質(zhì)體轉(zhuǎn)染法將構(gòu)建的ARC-miRNA及對照no-targe-miRNA質(zhì)粒轉(zhuǎn)染高表達(dá)ARC的CNE-2細(xì)胞；將pEZ-M03-ARC及對照pReceiver-M03CT質(zhì)粒穩(wěn)定轉(zhuǎn)染低表達(dá)ARC的6-10B細(xì)胞。通過熒光顯微鏡觀察質(zhì)粒轉(zhuǎn)染效率,CCK-8法檢測細(xì)胞生存率,熒光定量PCR、Western blot法檢測ARC表達(dá)效果,通過流式細(xì)胞儀檢測ARC對細(xì)胞周期的影響。 結(jié)果 1)三種miRNA干擾質(zhì)粒ARC-miRNA-1、ARC-miRNA-2、 ARC-miRNA-3及陰性對照質(zhì)粒轉(zhuǎn)染高表達(dá)ARC的CNE-2細(xì)胞,穩(wěn)定轉(zhuǎn)染后,熒光顯微鏡下綠色熒光表達(dá)水平均超過90%,細(xì)胞存活率分別為ARC-miRNA-1(98.4±1.3%)、ARC-miRNA-2(97.7±0.9%)、 ARC-miRNA-3(97.1±1.1%)、陰性對照組(98.1±1.6%)。pEZ-M03-ARC質(zhì)粒及陰性對照質(zhì)粒轉(zhuǎn)染低表達(dá)ARC的6-10B細(xì)胞,穩(wěn)定轉(zhuǎn)染后,熒光顯微鏡下綠色熒光表達(dá)水平均超過90%,細(xì)胞存活率分別為pEZ-M03-ARC (98.9±0.8%),陰性對照組(98.3±1.2%)。 2)熒光定量PCR檢測各組ARC mRNA表達(dá)水平。設(shè)定空白對照組ARC mRNA的表達(dá)量為1, ARC-miRNA-1、ARC-miRNA-2、 ARC-miRNA-3及陰性對照組CNE-2細(xì)胞中ARC mRNA的相對表達(dá)量分別為0.833±0.126、0.749±0.117、0.244±0.059、0.975±0.083；pEZ-M03-ARC及對照組6-10B細(xì)胞中ARC mRNA的相對表達(dá)量分別為2.371士0.330、1.149±0.166。 3)用Western blot檢測各組ARC基因蛋白表達(dá)水平,定空白對照組ARC蛋白的表達(dá)量為1,ARC-miRNA-1、ARC-miRNA-2、 ARC-miRNA-3及陰性對照組CNE-2細(xì)胞中ARC蛋白的相對表達(dá)量分別為0.875±0.144、0.796±0.122、0.271±0.082、0.989±0.130；pEZ-M03-ARC及對照組6-10B細(xì)胞中ARC蛋白的相對表達(dá)量分別為2.189±0.274、1.157±0.244。 4)穩(wěn)定轉(zhuǎn)染后檢測顯示ARC-miRNA-3質(zhì)粒為RNA干擾效果最強的質(zhì)粒。其在CNE-2細(xì)胞mRNA水平沉默效率為77.03%；蛋白水平沉默效率為72.6%。穩(wěn)定轉(zhuǎn)染pEZ-M03-ARC質(zhì)粒后檢測顯示：其在6-10B細(xì)胞mRNA水平的表達(dá)較對照組上調(diào)2.06倍；蛋白水平比對照組上調(diào)1.89倍。 5) ARC-miRNA-3CNE-2細(xì)胞的G0-G1、G2-M、S期的比例分別為56.5±4.7%、8.6±2.1%、34.8±2.5%,對照組CNE-2細(xì)胞的G0-G1、 G2-M、S期的比例分別為57.4±4.6%、9.1±2.3%、33.5士2.6%,兩組之間無明顯統(tǒng)計學(xué)差異(P0.05)；pEZ-M03-ARC6-10B細(xì)胞的G0-G1、 G2-M、S期的比例分別為61.5±3.3%、9.2±1.7%、29.3±2.3%,對照組6-10B細(xì)胞的G2-M、S期的比例分別為61.3±3.5%、10.3±2.1%、28.4±2.2%,兩組之間無明顯統(tǒng)計學(xué)差異(P0.05)。 結(jié)論成功構(gòu)建并篩選出miRNA干擾質(zhì)粒ARC-miRNA-3,并培養(yǎng)出穩(wěn)定沉默ARC表達(dá)的鼻咽癌細(xì)胞株；成功構(gòu)建并篩選出基因克隆質(zhì)粒pEZ-M03-ARC,并培養(yǎng)出穩(wěn)定過表達(dá)ARC的鼻咽癌細(xì)胞株。ARC的表達(dá)對鼻咽癌細(xì)胞的生長增殖及細(xì)胞周期無明顯影響。 目的探討沉默或過表達(dá)ARC后對鼻咽癌細(xì)胞放射敏感性的影響 方法沉默或過表達(dá)ARC的鼻咽癌細(xì)胞及對照組細(xì)胞分別接受不同劑量梯度的X線照射后,CCK-8法檢測細(xì)胞生存率,Annexin V法檢測細(xì)胞凋亡,Caspase3/8活性檢測試劑盒檢測Caspase3/8酶活性。 結(jié)果 1)沉默ARC表達(dá)的ARC-miRNA-3及對照組CNE-2細(xì)胞接受OGy、2Gy、4Gy、6Gy、8Gy、10Gy不同劑量照射后CCK-8法檢測兩組細(xì)胞存活率。ARC-miRNA-3細(xì)胞的細(xì)胞存活率明顯低于對照組細(xì)胞,實驗組細(xì)胞存活曲線明顯低于對照組,2Gy、4Gy、6Gy、8Gy、10Gy ARC-miRNA-3組細(xì)胞生存率分別為0.907±0.014、0.814±0.086、0.694±0.069、0.540±0.066、0.387±0.043；對照組細(xì)胞生存率分別為0.985±0.018、0.966±0.095、0.870±0.089、0.735±0.063、0.594±0.053(P0.001)；8Gy X線照射后,Annexin V法檢測ARC-miRNA-3細(xì)胞凋亡比例為52.2±3.4%明顯高于對照組細(xì)胞36.1±2.2%(P0.001)。 2)pEZ-M03-ARC細(xì)胞的細(xì)胞存活率明顯高于對照組細(xì)胞,實驗組細(xì)胞存活曲線明顯高于對照組,2Gy、4Gy、6Gy、8Gy、10Gy ARC-miRNA-3組細(xì)胞生存率分別為0.977±0.010、0.954±0.015、0.914±0.029、0.847±0.027、0.743±0.034；對照組細(xì)胞生存率分別為0.974±0.013、0.936±0.035、0.863±0.029、0.765±0.030、0.616±0.052(P0.001)；8Gy X線照射后,Annexin V法檢測細(xì)胞凋亡比例為21.2±2.4%明顯低于對照組細(xì)胞33.7±2.5%。 3)8Gy X線照射后,定義照組含量為1,ARC-miRNA-3CNE-2細(xì)胞的活性caspase-3含量為2.33±0.26,明顯高于對照組；ARC-miRNA-3CNE-2細(xì)胞的活性caspase-8含量為1.65±0.17,明顯高于對照組(P0.001)。 4)8Gy X線照射后,定義照組含量為1,pEZ-M03-ARC6-10B細(xì)胞的活性caspase-3含量為0.62±0.14,明顯低于對照組；pEZ-M03-ARC6-10B細(xì)胞的活性caspase-8含量為0.73±0.11,明顯低于對照組(P0.001)。 結(jié)論ARC有增強鼻咽癌細(xì)胞放射抵抗性的功能,并可能是通過抑制內(nèi)源性和外源性的凋亡通路來實現(xiàn)的,ARC有望成為增加鼻咽癌放射敏感性的分子治療靶點。 目的探討沉默或過表達(dá)ARC后對鼻咽癌化療藥物敏感性的影響 方法沉默或過表達(dá)ARC的鼻咽癌細(xì)胞及對照組細(xì)胞分別接受以不同濃度梯度的紫杉醇或順鉑處理48小時后,使用CCK-8法檢測細(xì)胞生長增殖情況。使用IC30的藥物濃度處理細(xì)胞,24小時后,使用Annxin V法檢測細(xì)胞凋亡情況,用Caspase3/8活性檢測試劑盒檢測Caspase3/8酶活性。 結(jié)果 1) ARC-miRNA-3細(xì)胞在順鉑中存活率明顯低于對照組細(xì)胞,實驗組細(xì)胞生存曲線明顯低于對照組(P0.001)。通過SPSS軟件計算,ARC-miRNA-3細(xì)胞順鉑作用IC50值為(1.995±0.327)×10-5明顯低于對照組IC50值(3.006±0.344)×10-5(P0.001)。Annexin V法檢測ARC-miRNA-3細(xì)胞凋亡比例為56.8±4.4%明顯高于對照組細(xì)胞33.02±3.7%(P0.001)。 2) pEZ-M03-ARC細(xì)胞在順鉑中細(xì)胞存活率明顯高于對照組細(xì)胞,實驗組細(xì)胞存活曲線明顯高于對照組(P0.001)；pEZ-M03-ARC細(xì)胞順鉑作用IC5o值為(1.000±0.244)×10-4明顯高于對照組IC50值(6.397±0.276)×10-5(P0.001)。Annexin V法檢測實驗組細(xì)胞凋亡比例為24.8±2.8%明顯低于對照組細(xì)胞35.1±2.6%(P0.001)。 3)順鉑處理后,ARC-miRNA-3CNE-2細(xì)胞的活性caspase-3含量與對照組的比例為3.49±0.31；ARC-miRNA-3CNE-2細(xì)胞的活性caspase-8含量與對照組的比例為2.17±0.20,均明顯高于對照組(P0.001)。 4)順鉑處理后,pEZ-M03-ARC6-10B細(xì)胞的活性caspase-3含量與對照組的比例為0.44±0.07; pEZ-M03-ARC6-1OB細(xì)胞的活性caspase-8含量與對照組的比例為0.69±0.12,均明顯低于對照組(P0.001)。 5)ARC-miRNA-3細(xì)胞及對照組細(xì)胞在紫杉醇中的存活率無明顯差別(P0.05); pEZ-M03-ARC細(xì)胞及對照組細(xì)胞在紫杉醇中的存活率無明顯差別(P0.05)。 結(jié)論ARC有增強鼻咽癌細(xì)胞化療抵抗的功能,并可能是通過抑制內(nèi)源性和外源性的凋亡通路來實現(xiàn)的,ARC有望成為增加鼻咽癌化療敏感性的分子治療靶點。
[Abstract]:Carcinogenesis is a complex process, which is caused by the interaction of oncogene activation, tumor suppressor genes and inactivation of cell death mechanism. The decrease of apoptosis is closely related to tumorigenesis, which suggests that genes involved in negative regulation of apoptosis may be carcinogenic, while genes involved in promoting apoptosis may be tumor suppression. Therefore, apoptosis-related proteins have always been a hot topic in tumor research. This article will discuss the relationship between a novel anti-apoptosis protein ARC and the occurrence and development of nasopharyngeal carcinoma, and its effect on radiotherapy and chemosensitivity of nasopharyngeal carcinoma.
Objective To investigate the relationship between the expression of ARC in nasopharyngeal carcinoma (NPC) and the clinical pathological parameters such as T stage, clinical stage and lymph node metastasis, and to detect the expression level and intracellular localization of ARC in NPC cell lines.
Methods Immunohistochemical method was used to detect paraffin-embedded tissues from 82 cases of nasopharyngeal carcinoma and 20 cases of chronic nasopharyngeal inflammation. The expression of.SPSS17.0 in nasopharyngeal carcinoma cell line was analyzed by statistical software.
Result
1) Immunohistochemical staining of nasopharyngeal carcinoma tissues and cells showed that ARC was expressed in cytoplasm and nucleus, most of which were expressed in cytoplasm.
2) The expression of ARC protein in nasopharyngeal carcinoma tissues was significantly higher than that in nasopharyngeal inflammatory tissues (P 0.001). Among nasopharyngeal carcinoma tissues, 44 were strongly positive (53.7%) 23 were moderately positive (28%) and 15 were weakly positive (18.3%).
3) The expression of ARC protein was correlated with the clinical stage and pathological type of NPC (P 0.001). The later the clinical stage, the higher the expression of ARC protein in the tissues with lower differentiation, but it was not correlated with age, sex and lymph node metastasis (P 0.05).
4) ARC was strongly expressed in nasopharyngeal carcinoma cell lines CNE-1, CNE-2, 5-8F, 6-10B, but weakly expressed in immortalized nasopharyngeal epithelial cells NP69. The relative expression levels of ARC were CNE-1 (4.78.47), CNE-2 (5.39.58), 5-8F (3.54.33), 6-10B (3.12.26), NP69 (1.67.24), respectively. The next time is CNE-2CNE-15-8F6-10B..
Conclusion These results suggest that ARC may play an important role in the occurrence and development of nasopharyngeal carcinoma. Detecting the expression of ARC may be helpful to predict the malignant degree of nasopharyngeal carcinoma.
Objective To screen stable transfected nasopharyngeal carcinoma cell lines by silencing and overexpressing ARC with RNA interference and gene cloning.
Methods RNA interference plasmids expressing microRNAs against ARC gene were constructed by pcDNA6.2-GW/EmGFP/microRNA vector, and plasmids expressing ARC gene were constructed by pEZ-M03 vector. ARC-microNA and control no-targe-microRNA plasmids were transfected into CNE-2 cells with high expression of ARC by liposome transfection, pEZ-M03-ARC and control pReceiver-M03CT plasmids were used. The plasmid transfection efficiency was observed by fluorescence microscope, the cell survival rate was detected by CCK-8, the expression of ARC was detected by fluorescence quantitative PCR and Western blot, and the effect of ARC on cell cycle was detected by flow cytometry.
Result
1) Three microRNAs interfered with the expression of ARC-microNA-1, ARC-microNA-2, ARC-microNA-3 and negative control plasmids in CNE-2 cells with high expression of ARC. After stable transfection, the expression of green fluorescence under fluorescence microscope exceeded 90%, and the cell survival rates were ARC-microNA-1 (98.4 (1.3%), ARC-microNA-2 (97.7 (0.9%), ARC-microNA-3 (97.1 (1) and negative control group (97.1). PEZ-M03-ARC plasmid and negative control plasmid were transfected into 6-10B cells with low expression of ARC. After stable transfection, the expression of green fluorescence under fluorescence microscope exceeded 90%. The survival rate of pEZ-M03-ARC cells was 98.9%(98.9%+0.8%) and that of negative control group was 98.3%+1.2%.
2) The expression of ARC mRNA in CNE-2 cells was detected by fluorescence quantitative PCR. The relative expression levels of ARC mRNA in blank control group were 1, ARC-microNA-1, ARC-microNA-2, ARC-microNA-3 and negative control group were 0.833 [0.126], 0.749 [0.117], 0.244 [0.059], 0.975 [0.083], pEZ-M03-ARC and 6-10B cells respectively. The relative expression amounts were 2.371 0.330,1.149 0.166..
3) Western blot was used to detect the expression level of ARC gene protein in each group. The relative expression levels of ARC protein in blank control group were 1, ARC-microNA-1, ARC-microNA-2, ARC-microNA-3 and negative control group CNE-2 cells were 0.875 (+ 0.144), 0.796 (+ 0.122), 0.271 (+ 0.082), 0.989 (+ 0.130), pEZ-M03-ARC and 6-10B cells respectively. The relative expression of protein was 2.189 + 0.274,1.157 + 0.244.
4) After stable transfection, ARC-microNA-3 plasmid was the most effective plasmid for RNA interference. Its silencing efficiency was 77.03% at mRNA level and 72.6% at protein level in CNE-2 cells. After stable transfection of pEZ-M03-ARC plasmid, the expression of ARC-microNA-3 in 6-10B cells was 2.06 times higher than that in control group. Up to 1.89 times.
5) The proportions of G0-G1, G2-M and S-phase of ARC-microNA-3CNE-2 cells were 56.5 (+ 4.7%), 8.6 (+ 2.1%) and 34.8 (+ 2.5%) respectively. The proportions of G0-G1, G2-M and S-phase of CNE-2 cells in control group were 57.4 (+ 4.6%), 9.1 (+ 2.3%) and 33.5 (+ 2.6%) respectively. There was no significant difference between the two groups (P 0.05). The proportions of G2-M and S-phase of 6-10B cells in the control group were 61.3 (+ 3.5%), 10.3 (+ 2.1%) and 28.4 (+ 2.2%) respectively. There was no significant difference between the two groups (P 0.05).
Conclusion The interfering plasmid ARC-microNA-3 was successfully constructed and screened, and the stable silencing cell line of nasopharyngeal carcinoma was cultured. The cloned plasmid pEZ-M03-ARC was successfully constructed and screened, and the stable overexpressing cell line of nasopharyngeal carcinoma was cultured.
Objective to investigate the effect of silencing or over expression of ARC on radiosensitivity of nasopharyngeal carcinoma cells.
Methods Nasopharyngeal carcinoma cells silenced or overexpressed ARC were irradiated with different dose gradient X-ray respectively. Cell survival rate was measured by CCK-8 method, apoptosis was detected by Annexin V method, and caspase 3/8 activity was detected by Caspase 3/8 activity assay kit.
Result
1) The survival rate of ARC-microNA-3 cells was measured by CCK-8 assay after different doses of OGy, 2Gy, 4Gy, 6Gy, 8Gy, 10Gy irradiation. The cell survival rate of ARC-microNA-3 cells was significantly lower than that of control cells, and the cell survival curve of experimental group was significantly lower than that of control group, 2Gy, 4Gy, 6Gy, 8Gy, 10Gy ARC-microNA-3 cells. The survival rates were 0.907 [0.907 [0.907 [0.017 [0.017 [0.014,0.814 [0.81 [0.086,0.694 [0.694 [0.069,0.699 9 9 [, 0.694 [, 0.540 [0.540 [0.066 [0.066 6 6 0.066 6, 0.066 0.060 [0.066 6 0.066 6, 0.387 [0.387 [0.387 [0.040.0403 [.043] 3] 3] respectively; the cell survival rates in control group were 0.985 5 5 5 In the meantime, it is necessary to study the relationship between the two. + 2.2% (P0.001).
2) The cell survival rate of pEZ-M03-ARC cells was significantly higher than that of the control group. The cell survival curves of the experimental group were significantly higher than that of the control group. The cell survival rates of the experimental group, 2Gy, 4Gy, 6Gy, 8Gy, 10Gy ARC-MiNA-3 groups were 0.977 [0.010], 0.954 [0.015], 0.914 [0.029], 0.847 [0.027], 0.743 [0.034], respectively. After 8 Gy X-ray irradiation, the percentage of apoptosis detected by Annexin V method was 21.2+2.4%, which was significantly lower than that of the control group (33.7+2.5%).
3) After 8 Gy X-ray irradiation, the content of active caspase-3 in ARC-microNA-3CNE-2 cells was 1 and 2.33+0.26 respectively, which was significantly higher than that in control group, and the content of active caspase-8 in ARC-microNA-3CNE-2 cells was 1.65+0.17, which was significantly higher than that in control group (P 0.001).
4) After 8 Gy X-ray irradiation, the content of active caspase-3 in pEZ-M03-ARC6-10B cells was 0.62+0.14, which was significantly lower than that in control group, and the content of active caspase-8 in pEZ-M03-ARC6-10B cells was 0.73+0.11, which was significantly lower than that in control group (P 0.001).
Conclusion ARC can enhance the radioresistance of nasopharyngeal carcinoma cells and may be achieved by inhibiting endogenous and exogenous apoptotic pathways. ARC may be a promising molecular therapeutic target for increasing radiosensitivity of nasopharyngeal carcinoma.
Objective to investigate the effect of silencing or over expression of ARC on chemosensitivity of nasopharyngeal carcinoma.
Methods Nasopharyngeal carcinoma cells silenced or overexpressed ARC were treated with paclitaxel or cisplatin at different concentration gradients for 48 hours. Cell growth and proliferation were detected by CCK-8 assay. Cells treated with IC30 were detected by Annxin V assay after 24 hours. Cell apoptosis was detected by Caspase 3/8 activity assay. The Caspase3/8 activity was detected by kit.
Result
1) The survival rate of ARC-microNA-3 cells in cisplatin was significantly lower than that of the control group, and the survival curve of the experimental group was significantly lower than that of the control group (P 0.001). The proportion of apoptotic cells was 56.8 + 4.4%, which was significantly higher than that of the control group (33.02 + 3.7%) (P0.001).
2) The cell survival rate of pEZ-M03-ARC cells in cisplatin was significantly higher than that of the control group, and the cell survival curve of the experimental group was significantly higher than that of the control group (P 0.001). The IC5o value of pEZ-M03-ARC cells treated by cisplatin was (1.000.244)*10-4, which was significantly higher than that of the control group (6.397.276)*10-5 (P 0.001). The apoptosis rate of the experimental group was 2.00% by Annexin V assay. 4.8 + 2.8% was significantly lower than that of the control group (35.1 + 2.6%) (P0.001).
3) After cisplatin treatment, the ratio of active caspase-3 in ARC-microNA-3CNE-2 cells to the control group was 3.49 [0.31], and the ratio of active caspase-8 in ARC-microNA-3CNE-2 cells to the control group was 2.17 [0.20], which was significantly higher than that in the control group (P 0.001).
4) After cisplatin treatment, the ratio of active caspase-3 in pEZ-M03-ARC6-10B cells to control group was 0.44.07, and the ratio of active caspase-8 in pEZ-M03-ARC6-1OB cells to control group was 0.69.12, which was significantly lower than that in control group (P 0.001).
5) There was no significant difference in the survival rate of ARC-microNA-3 cells and control cells in paclitaxel (P 0.05); there was no significant difference in the survival rate of pEZ-M03-ARC cells and control cells in paclitaxel (P 0.05).
Conclusion ARC can enhance the chemotherapeutic resistance of nasopharyngeal carcinoma cells and may be achieved by inhibiting endogenous and exogenous apoptotic pathways. ARC is expected to be a molecular therapeutic target for increasing the chemotherapeutic sensitivity of nasopharyngeal carcinoma cells.
【學(xué)位授予單位】：中南大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2012
【分類號】：R739.63

【共引文獻(xiàn)】

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2 張曉偉;DNA甲基化在鼻咽癌紫杉醇耐藥形成及逆轉(zhuǎn)耐藥機制中的初步研究[D];中南大學(xué);2012年

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