發(fā)布時(shí)間：2018-05-14 23:19

  本文選題：鼻咽腫瘤 + 環(huán)氧合酶-2��； 參考：《南方醫(yī)科大學(xué)》2011年碩士論文

【摘要】：研究背景及目的： 鼻咽癌(nasopharyngeal carcinoma, NPC)是我國常見的惡性腫瘤之一,其發(fā)病以明顯的區(qū)域分布、強(qiáng)烈的轉(zhuǎn)移傾向、與EB病毒的密切關(guān)系為顯著特征。放射治療是鼻咽癌目前公認(rèn)和有效的治療手段,但因其嚴(yán)重的并發(fā)癥及后遺癥,治療效果并不穩(wěn)定,5年總生存率始終徘徊在50%左右。因此,探究鼻咽癌發(fā)病的分子機(jī)制,尋找新的治療靶點(diǎn),提高現(xiàn)有治療手段的療效,是當(dāng)代腫瘤學(xué)研究的一項(xiàng)迫切任務(wù)。 COX-2是環(huán)氧合酶兩種同工酶的其中一種,在體內(nèi)呈誘導(dǎo)性表達(dá),在大多數(shù)正常組織中難以發(fā)現(xiàn)。當(dāng)受到一些刺激因子如生長因子、細(xì)胞因子及促癌劑的刺激時(shí),其表達(dá)量會成倍增加。抑制COX-2的表達(dá)可明顯增加化療及放療對鼻咽癌細(xì)胞的殺傷能力。 shRNAmir(microRNA-adapted,shRNA)是在microRNA結(jié)構(gòu)基礎(chǔ)上改造出的發(fā)夾狀小干擾RNA,利用shRNAmir表達(dá)載體,在細(xì)胞內(nèi)轉(zhuǎn)錄出與miRNA相似的shRNAmir,比傳統(tǒng)的shRNA更加安全有效,是人工誘導(dǎo)RNA干擾沉默基因表達(dá)的最新技術(shù)。 本課題前期已完成如下研究工作：應(yīng)用RNA干擾技術(shù)沉默COX-2的表達(dá),使得鼻咽癌細(xì)胞周期出現(xiàn)G0/G1期阻滯,且增殖能力明顯下降；應(yīng)用基于miR-155結(jié)構(gòu)的RNA干擾技術(shù),成功構(gòu)建了Anti-COX-2 shRNAmir(?)慢病毒表達(dá)載體,經(jīng)慢病毒感染48-72h后,陽性C666-1細(xì)胞發(fā)出綠色螢光,感染效率最高達(dá)90.5%,經(jīng)流式細(xì)胞儀分選出帶熒光的細(xì)胞經(jīng)連續(xù)傳代3個(gè)月后,帶光率仍超99%,RT-PCR幾乎檢測不到COX-2基因mRNA的表達(dá),獲得了穩(wěn)定抑制COX-2表達(dá)的鼻咽癌細(xì)胞亞系,證明shRNAmir慢病毒載體具備獲得穩(wěn)定持久的RNA干擾作用,在轉(zhuǎn)錄后水平沉默COX-2基因,達(dá)到了“基因敲除”的效果,遠(yuǎn)優(yōu)于Anti-COX-2 siRNA的干擾效果,應(yīng)用于鼻咽癌分子機(jī)制研究,為下一步的分子靶向治療和基因-放射聯(lián)合治療奠定基礎(chǔ)。 在前期研究工作的基礎(chǔ)上,本研究進(jìn)一步探討COX-2基因沉默后對鼻咽癌細(xì)胞生長、轉(zhuǎn)移及對放化療敏感性的影響,為鼻咽癌患者的綜合治療提供理論依據(jù)。 方法 1.檢測COX-2沉默后對鼻咽癌細(xì)胞生長特性的影響 2×103個(gè)指數(shù)生長期細(xì)胞接種于96孔板中,每孔細(xì)胞懸液量為100ul,置于培養(yǎng)箱中分別培養(yǎng)1、2、3、4、5天,向每孔加入10ul的CCK-8溶液,將培養(yǎng)板置于培養(yǎng)箱中孵育1h,用酶標(biāo)儀測定在450nm處的OD值,以相對應(yīng)OD值表示細(xì)胞增殖能力大小。每組設(shè)6個(gè)重復(fù)孔,取平均值,繪制體外生長曲線。 2.劃痕實(shí)驗(yàn) 2×105個(gè)細(xì)胞接種于6孔板,每組細(xì)胞作3個(gè)復(fù)孔,待細(xì)胞長至約90%匯合時(shí),吸干培養(yǎng)基,用200ul tip頭垂直在培養(yǎng)孔中部輕輕劃過,力度以刮落細(xì)胞而不在培養(yǎng)板上留痕為準(zhǔn)。PBS洗2遍以沖去脫落細(xì)胞,加入培養(yǎng)基后繼續(xù)培養(yǎng)。每個(gè)孔隨機(jī)選取9個(gè)有劃痕穿過的視野,分別觀察0h,6h,12h,24h時(shí)細(xì)胞細(xì)胞劃痕處細(xì)胞運(yùn)動(dòng)變化,并拍照取樣,應(yīng)用Image J軟件測劃痕相對寬度,繪制遷移力量值曲線,獨(dú)立實(shí)驗(yàn)重復(fù)3次。 3.檢測COX-2沉默后鼻咽癌細(xì)胞對順鉑的敏感性變化 兩組細(xì)胞分別按5000個(gè)/孔加入96孔板中,每孔加入培養(yǎng)基100ul,培養(yǎng)24h后棄去培養(yǎng)基,加入含有不同濃度順鉑的培養(yǎng)基,使其終濃度分別為0.5、1、2、4、8、16、32ug/ml,每組3個(gè)復(fù)孔,并設(shè)不接種細(xì)胞的空白培養(yǎng)基調(diào)零組和加入等體積溶劑的對照組,作用24h后加入10ul CCK-8試劑,置于培養(yǎng)箱中孵育1h后,利用酶標(biāo)儀測各孔450nm處的吸光度A450,計(jì)算細(xì)胞生長的抑制率,并應(yīng)用SPSS13.0軟件計(jì)算半數(shù)抑制濃度。 4.細(xì)胞周期實(shí)驗(yàn) 指數(shù)生長期細(xì)胞在含有6ug/ml的順鉑或不含順鉑的培養(yǎng)基中培養(yǎng)24h,或經(jīng)2Gy照射或0Gy假照射24及48h后,以不含EDTA的胰酶消化后收集細(xì)胞,加入70%冰乙醇中4℃過夜,PBS洗滌2遍后,避光加入含有50ug/ml的RNA酶和50ul/ml PI的染色液中,室溫染色30min后,流式細(xì)胞儀檢測細(xì)胞周期各時(shí)相所占百分比。 5.克隆形成實(shí)驗(yàn) 兩組細(xì)胞按預(yù)定數(shù)量接種于6孔板中,每組均設(shè)0、1、2、4、6、8Gy六個(gè)劑量組,每一劑量組3個(gè)復(fù)孔。照射后的細(xì)胞置于37℃、5%CO2培養(yǎng)箱內(nèi)培養(yǎng)。當(dāng)出現(xiàn)肉眼可見克隆時(shí),終止培養(yǎng),甲醇固定,1%結(jié)晶紫乙醇溶液染色。顯微鏡下計(jì)數(shù)含50個(gè)細(xì)胞以上的克隆數(shù),計(jì)算克隆形成率和存活分?jǐn)?shù)。以3次照射的存活分?jǐn)?shù)均值進(jìn)行分析,運(yùn)用GraphPad Prism 5.0軟件進(jìn)行L-Q模型和多靶單擊模型曲線擬合,繪制劑量存活曲線,并計(jì)算L-Q模型參數(shù)α、β、α/β、SF2和多靶單擊模型參數(shù)D0、Dq、和N,其中l(wèi)ogN=Dq/D0,計(jì)算放射增敏比。 6.體內(nèi)放療敏感性實(shí)驗(yàn) 雌性BALB/c裸鼠,4-6周齡,18～22g, SPF條件下飼養(yǎng)。將兩組細(xì)胞分別制成單細(xì)胞懸液,用無血清1640培養(yǎng)基調(diào)節(jié)細(xì)胞濃度至1×106/ml。在裸鼠右后肢皮下接種細(xì)胞懸液0.1ml,實(shí)驗(yàn)中每2天用游標(biāo)卡尺測量腫瘤的最大徑和與之垂直的橫徑,腫瘤體積(V)=(長徑×短徑2)/2。10-14d后,當(dāng)腫瘤體積達(dá)到約200mm3時(shí),將荷瘤裸鼠分為4組：①COX-2(+)對照組,無任何處理；②COX-2(-)對照組,無任何處理；③COX-2(+)實(shí)驗(yàn)組,給予10GyX線照射；④COX-2(-)實(shí)驗(yàn)組,給予10Gy X線照射。每組6只,2周后斷頸處死裸鼠,計(jì)算抑瘤率。 7.統(tǒng)計(jì)學(xué)分析 采用SPSS13.0軟件進(jìn)行統(tǒng)計(jì)分析,實(shí)驗(yàn)數(shù)據(jù)以均數(shù)±標(biāo)準(zhǔn)差表示,兩樣本均數(shù)的比較才采用完全隨機(jī)設(shè)計(jì)資料的方差分析(One-Way ANOVA)或兩獨(dú)立樣本的t檢驗(yàn)(Independent-Sample T Test)。增殖實(shí)驗(yàn)和劃痕實(shí)驗(yàn)采用析因設(shè)計(jì)的方差分析,多重比較采用SNK法(Student-Neuman-Keuls), P0.05表示差異有統(tǒng)計(jì)學(xué)意義。 結(jié)果： 1.COX-2基因沉默后對鼻咽癌細(xì)胞增殖和體外遷移能力的影響 Anti-COX-2 C666-1和Anti-GL-2 C666-1細(xì)胞,接種96孔板連續(xù)觀察5天,依據(jù)450nm處OD值繪制生長曲線,析因分析結(jié)果表明,兩種細(xì)胞增殖差異有統(tǒng)計(jì)學(xué)意義(F=5.927,P=0.019),但細(xì)胞組別與天數(shù)之間無交互效應(yīng)(F=1.264,P=0.296),說明COX-2基因沉默后細(xì)胞增殖能力有所下降,但兩組細(xì)胞增殖速度隨時(shí)間變化趨勢相同。 細(xì)胞劃痕實(shí)驗(yàn)檢測COX-2穩(wěn)定沉默后對鼻咽癌細(xì)胞C666-1體外遷移能力的影響,析因設(shè)計(jì)的方差分析結(jié)果顯示,隨著培養(yǎng)時(shí)間的延長,兩組間劃痕寬度差異有統(tǒng)計(jì)學(xué)意義(F=157.01,P0.001),細(xì)胞組別與時(shí)間之間有交互效應(yīng)(F=15.73,P0.001),說明隨著時(shí)間的變化,兩組細(xì)胞體外遷移距離的變化趨勢不同,Anti-COX-2 C666-1細(xì)胞遷移能力較Anti-GL-2 C666-1有所減弱。 2.COX-2沉默后鼻咽癌C666-1細(xì)胞對順鉑敏感性的變化 析因分析結(jié)果顯示,不同濃度順鉑作用24h后,兩組細(xì)胞增殖抑制率差異有統(tǒng)計(jì)學(xué)意義(F=22.219,P0.001),經(jīng)計(jì)算得出Anti-COX-2 C666-1和Anti-GL-2C666-1細(xì)胞對順鉑的IC50分別為3.682±0.980和8.819±2.354,兩組間IC50差異有統(tǒng)計(jì)學(xué)意義(t=-3.489,P=0.025)。 3.COX-2沉默后對順鉑引起細(xì)胞周期變化的影響 流式細(xì)胞儀分析結(jié)果表明,加順鉑前Anti-COX-2 C666-1和Anti-GL-2C666-1細(xì)胞周期中,G0/G1期細(xì)胞比例分別為70.260%和60.353%(t=-2.726,P=0.053),S期細(xì)胞比例分別為24.270%和32.180%(t=-2.033,P=0.112),差異均無統(tǒng)計(jì)學(xué)意義,G2/M期細(xì)胞比例差別有統(tǒng)計(jì)學(xué)意義,分別為5.470%和7.500%(t=-3.954, P=0.017)。6ug/ml的順鉑作用24h后,兩組細(xì)胞S期與G2/M期比例均升高,兩組均出現(xiàn)S期和G2/M期阻滯,S期比例升高幅度分別為24.967%和32.200%,兩組間差異無統(tǒng)計(jì)學(xué)意義(t=--1.636,P=0.177),G2/M期比例升高幅度分別為0.500%和7.633%,兩組間差異有統(tǒng)計(jì)學(xué)意義(t=-7.501,P=0.002)。COX-2沉默后G2/M期升高比例減少。但順鉑作用前后均未出現(xiàn)明顯的凋亡峰,說明COX-2沉默后降低了順鉑引起的細(xì)胞G2/M期阻滯,但對細(xì)胞凋亡無明顯影響。 4.COX-2沉默后對鼻咽癌C666-1細(xì)胞體外放療敏感性的影響 COX-2沉默前后Do值分別為2.0157±0.0515、1.6333±0.0751,Dq值分別為0.8037±0.0487、0.5643±0.0399,SF2分別為0.5551±0.0375、0.3961±0.0160,α/β分別為6.5356±0.8461、9.4231±0.9599,兩組間D0、Dq、F2、α/β值差異均有統(tǒng)計(jì)學(xué)意義,放射增敏比SER=1.4014。 5.COX-2沉默后對輻射引起細(xì)胞周期的影響 流式細(xì)胞儀分析結(jié)果表明,照射前Anti-GL-2 C666-1和Anti-COX-2 C666-1細(xì)胞周期中,G0/G1以及S期細(xì)胞比例差異均無統(tǒng)計(jì)學(xué)意義,但G2/M期細(xì)胞比例差別有統(tǒng)計(jì)學(xué)意義,分別為7.500%和5.470%(t=-3.952,P=0.017)。照射2Gy后24h,G2期細(xì)胞比例均增高,分別為45.833%和23.900%(t=11.999,P0.001),兩組細(xì)胞均出現(xiàn)G2/M期阻滯,但Anti-GL-2 C666-1細(xì)胞G2/M期升高比例更大,為38.333%,而Anti-COX-2 C666-2細(xì)胞G2/M期比例升高18.430%(t=9.184,P=0.001)。照射2Gy后48h,兩組細(xì)胞G2/M期的比例分別為24.397%和12.393%(t=4.646,P=0.01),較照射前分別增加了17.357%和6.923%(t=3.320,P=0.029)。但照射前后均未出現(xiàn)反映細(xì)胞凋亡的亞G1期(sub-G1 phase),說明COX-2沉默后顯著降低了輻射引起的細(xì)胞G2/M期阻滯,但對細(xì)胞凋亡無明顯影響。 6.COX-2沉默后對體內(nèi)荷瘤裸鼠輻射敏感性的影響 未放療組,Anti-GL-2和Anti-COX-2兩組裸鼠移植瘤生長曲線幾乎重合,COX-2沉默前后對皮下移植瘤的生長情況差異無統(tǒng)計(jì)學(xué)意義(F=3.061,P=0.084),加入10Gy單純放療后,Anti-GL-2和Anti-COX-2兩組的抑瘤率分別為32.72%和51.76%,兩組間皮下移植瘤的生長情況差異有統(tǒng)計(jì)學(xué)意義(F=63.468,P0.001)。 結(jié)論： 1.COX-2沉默后降低了鼻咽癌細(xì)胞的增殖及體外遷移能力； 2.COX-2沉默后增強(qiáng)順鉑對鼻咽癌C666-1細(xì)胞的敏感性,降低了順鉑引起的細(xì)胞G2/M期阻滯； 3.COX-2沉默后增強(qiáng)鼻咽癌體內(nèi)外放療的敏感性,降低了輻射引起的細(xì)胞G2/M期阻滯。
[Abstract]:Background and purpose of the study :

Nasopharyngeal carcinoma ( NPC ) is one of the most common malignant tumors in China . Its pathogenesis is characterized by obvious regional distribution , strong metastasis tendency and close relationship with EB virus . Radiotherapy is the most commonly accepted and effective treatment method for nasopharyngeal carcinoma .

COX - 2 is one of the two isozymes of cyclooxygenase , which is inducible in vivo . It is difficult to find in most normal tissues . When stimulated by some stimulating factors such as growth factors , cytokines and cancer promoting agents , the expression of COX - 2 can be increased . The inhibition of COX - 2 expression can significantly increase the killing ability of chemotherapy and radiotherapy for nasopharyngeal carcinoma cells .

shRNAmir ( microRNA - adapted , shRNA ) is a hairpin - shaped small interfering RNA modified on the basis of microRNA structure , and shRNAmir similar to miRNA is transcribed in the cell by using the shRNAmir expression vector , and is more safe and effective than the conventional shRNA , and is an up - to - date technology for artificially inducing RNA interference silencing gene expression .

In the early stage of the project , we have completed the following research : silencing the expression of COX - 2 by using RNA interference technique , so that the cell cycle of nasopharyngeal carcinoma appears G0 / G1 phase arrest , and the proliferation ability is obviously decreased ;
The anti - COX - 2 shRNAmir ( ? ) lentivirus expression vector was successfully constructed by using RNA interference technique based on the structure of miR - 155 . After 48 to 72 hours of lentivirus infection , the positive C666 - 1 cells emitted green fluorescent light . After three months of continuous passage , the expression of COX - 2 mRNA was detected .

Based on previous studies , this study further explores the effects of COX - 2 gene silencing on the growth , metastasis and sensitivity of nasopharyngeal carcinoma cells ( NPC ) cells and provides a theoretical basis for the comprehensive treatment of nasopharyngeal carcinoma patients .

method

5 . Effect of COX - 2 silencing on cell cycle induced by radiation

2 脳 103 exponential growth cells were inoculated into 96 well plates , the cell suspension volume was 100ul , cultured in incubator for 1 , 2 , 3 , 4 , 5 days , and 10 ul of CCK - 8 solution was added to each well . The OD value at 450nm was measured by microplate reader . Six replicate wells were set in each group , and the average value was taken to plot the growth curve in vitro .

2 . Scratch test

2 脳 105 cells were seeded on 6 - well plates . Each group of cells was divided into three complex wells . When the cells grew to about 90 % confluence , the culture medium was washed twice with 200 ul tip . The cells were washed twice to remove the cells . After addition of the culture medium , the cells were washed twice to remove the cells . Then the cells were taken for sampling . The relative widths of the scratches were measured with the image J software . The displacement force curve was drawn and the independent experiment was repeated three times .

3 . Sensitivity changes of nasopharyngeal carcinoma cells to cisplatin after COX - 2 silencing

Two groups of cells were added into 96 well plates according to 5000 cells / well , medium 100ul was added into each well , the culture medium was discarded after 24 h , the final concentration was 0.5 , 1 , 2 , 4 , 8 , 16 , 32 ug / ml , and the final concentration was 0.5 , 1 , 2 , 4 , 8 , 16 , 32 ug / ml . After 24 h , 10 ul of CCK - 8 reagent was added . After incubation for 1h , the inhibitory rate of cell growth was calculated , and half of the inhibition concentration was calculated by SPSS 13.0 software .

4 . Cell cycle experiment

The cells were cultured for 24 h in a medium containing 6 ug / ml of cis - platinum or no cisplatin , or 24 and 48 hours after irradiation with 2 Gy or 0 Gy of sham irradiation . After 2 times of washing with PBS , the cells were added to the staining solution containing 50 ug / ml RNase and 50 ul / ml PI . After 30 min at room temperature , the percentage of each phase of cell cycle was detected by flow cytometry .

5 . Cloning formation experiment

Two groups of cells were inoculated into 6 - well plates according to a predetermined number . Each group was set with 6 dose groups of 0 , 1 , 2 , 4 , 6 and 8 Gy . After irradiation , the cells were cultured in a 5 % CO2 incubator . The results were analyzed by using GraphPad Prism 5.0 software . The parameters of L - Q model , 尾 , 偽 / 尾 , SF2 and multi - target were calculated . Then , the parameters D0 , Dq , and N were calculated .

6 . Radiosensitivity test in vivo

Female BALB / c nude mice , 4 - 6 week old , 18 - 22 g , SPF condition were fed . Two groups of cells were respectively made into single cell suspension , the cell concentration was adjusted to 1 脳 106 / ml with serum - free 1640 medium . The tumor volume ( V ) = ( long diameter 脳 short diameter 2 ) / 2.10 - 14d was measured by vernier caliper in the right hind limb of nude mice .
( 2 ) COX - 2 ( - ) control group without any treatment ;
( 3 ) COX - 2 ( + ) experimental group was given 10 Gy X - ray irradiation ;
( 4 ) COX - 2 ( - ) experimental group was given 10 Gy X - ray irradiation . 6 rats in each group were sacrificed 2 weeks later , the nude mice were sacrificed and the tumor inhibition rate was calculated .

7 . Statistical Analysis

Statistical analysis was carried out with SPSS 13.0 software , and the experimental data was expressed by mean 鹵 SD , and the comparison of the two samples was conducted using One - Way ANOVA or Independent - Sample T Test of two independent samples . The results showed that the difference was statistically significant by using SNK method ( Student - Neuman - Keeper ) and P0.05 .

Results :

1 . Effect of COX - 2 gene silencing on proliferation and in vitro migration of nasopharyngeal carcinoma cells

Anti - COX - 2 C666 - 1 and Anti - GL - 2 C666 - 1 cells were inoculated with 96 well plates for 5 days . The growth curve was plotted on the basis of OD value at 450nm . The results showed that there was no interaction between the two groups ( F = 5.927 , P = 0.019 ) , but there was no interaction between cell group and days ( F = 1.264 , P = 0.296 ) .

The effect of COX - 2 stable silencing on the in vitro migration of C666 - 1 in nasopharyngeal carcinoma cells was detected by cell scratch test . The results of variance analysis showed that the difference between the two groups had statistical significance ( F = 157.01 , P0.001 ) , and there was an interaction effect between the two groups ( F = 15.73 , P0.001 ) . The migration distance of the two groups was different with time . Anti - COX - 2 C666 - 1 cells migrated more than Anti - GL - 2 C666 - 1 .

2 . Changes of the sensitivity of C666 - 1 cells to cisplatin after COX - 2 silencing

The IC50 of anti - COX - 2 C666 - 1 and Anti - GL - 2C666 - 1 cells on cisplatin was 3.682 鹵 0.980 and 8.819 鹵 2.354 , respectively , and the IC50 values of anti - COX - 2 C666 - 1 and Anti - GL - 2C666 - 1 cells were 3.682 鹵 0.980 and 8.819 鹵 2.354 respectively .

3 . Effect of COX - 2 on Cell Cycle Changes Induced by Cisplatin

The results of flow cytometry showed that the percentage of G0 / G1 phase cells was 70.260 % and 60.353 % ( t = - 2.726 , P = 0.053 ) , and the percentage of S - phase cells was 24.27 % and 32.180 % ( t = - 3.954 , P = 0.112 ) , respectively . The percentage of S - phase and G2 / M phase increased . There was no significant difference between the two groups ( t = - 1.636 , P = 0.177 ) . The amplitude of G2 / M phase increased by 0.500 % and 7.633 % , respectively . There were no obvious apoptotic peaks in the G2 / M phase after COX - 2 silencing , but there were no obvious apoptotic peaks before and after the action of cisplatin , suggesting that the cell G2 / M phase block induced by cisplatin was decreased after COX - 2 silencing , but there was no obvious effect on apoptosis .

4 . Effect of COX - 2 silencing on the radiosensitivity of C666 - 1 cell in nasopharyngeal carcinoma

The values of D0 , Dq , F2 and 偽 / 尾 in the two groups were 6.5356 鹵 0.0375 , 0.3961 鹵 0.0160 , 6.5356 鹵 0.861 , 9.4231 鹵 0.9599 , respectively .

1 . Effect of COX - 2 gene silencing on the growth characteristics of nasopharyngeal carcinoma cells

The results of flow cytometry showed that there was no significant difference between G0 / G1 and S phase in the cell cycle of Anti - GL - 2 C666 - 1 and Anti - COX - 2 C666 - 1 , but the percentage of G2 / M cells was 7.500 % and 5.470 % , respectively ( t = - 3.952 , P = 0 . 017 ) . The G2 / M phase of anti - GL - 2 C666 - 1 cells increased by 18.430 % ( t = 9.184 , P = 0.001 ) . The G2 / M phase of anti - GL - 2 C666 - 1 cells increased by 18.430 % ( t = 9.184 , P = 0.001 ) . The G2 / M phase of the two groups were 24.397 % and 12.393 % ( t = 4.646 , P = 0.01 ) after 2Gy irradiation , 17.357 % and 6.923 % respectively before irradiation ( t = 3.320 , P = 0.029 ) . However , there were no sub - G1 phase reflecting apoptosis before and after irradiation , indicating that COX - 2 silencing significantly reduced the G2 / M phase block induced by radiation , but had no obvious effect on apoptosis .

6 . Effect of COX - 2 silencing on radiation sensitivity of nude mice in vivo

There was no significant difference in growth curve between the two groups ( F = 3.61 , P = 0.084 ) , and the tumor inhibition rates of anti - GL - 2 and Anti - COX - 2 groups were 32.72 % and 51.76 % , respectively .

Conclusion :

1 . COX - 2 silencing reduces the proliferation and in vitro migration ability of nasopharyngeal carcinoma cells .


2 . The sensitivity of cisplatin to nasopharyngeal carcinoma ( C666 - 1 ) was enhanced after COX - 2 silencing , and cell G2 / M arrest induced by cisplatin was reduced .


3 . After the COX - 2 silencing , the sensitivity of external radiotherapy in nasopharyngeal carcinoma was enhanced , and the G2 / M phase block of the cells induced by radiation was reduced .

【學(xué)位授予單位】：南方醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2011
【分類號】：R739.63

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