【摘要】：目的研究FANCD2 sh RNA干擾對(duì)人頭頸鱗癌(head and neck squamous cell carcinoma,HNSCC)細(xì)胞SIHN-005A化療敏感度的影響,并初步探討其可能機(jī)制。方法應(yīng)用前期實(shí)驗(yàn)獲得的FANCD2基因沉默效應(yīng)穩(wěn)定的人HNSCC SIHN-005A細(xì)胞,以嘌呤霉素持續(xù)篩選;Western blot檢測(cè)沉默效應(yīng);CCK-8法檢測(cè)順鉑作用后細(xì)胞增殖抑制率;Hoechst法檢測(cè)細(xì)胞凋亡;Western blot檢測(cè)Cyclin D1、Bax/Bcl-2、HIF-1α蛋白的表達(dá)。結(jié)果實(shí)驗(yàn)組細(xì)胞FANCD2蛋白表達(dá)明顯下降,FANCD2蛋白沉默效率達(dá)60.5%。實(shí)驗(yàn)組細(xì)胞增殖抑制率明顯高于陰性對(duì)照組和空白對(duì)照組,且具有濃度及時(shí)間依賴性,實(shí)驗(yàn)組細(xì)胞對(duì)CDDP的IC50值明顯低于陰性對(duì)照組和空白對(duì)照組。FANCD2基因沉默后實(shí)驗(yàn)組細(xì)胞凋亡率明顯增高。加用CDDP后,Cyclin D1、Bcl-2、HIF-1α蛋白表達(dá)水平較加用CDDP前明顯下調(diào),Bax蛋白明顯增強(qiáng)。結(jié)論沉默F(xiàn)ANCD2可增加SIHN-005A細(xì)胞對(duì)化療藥物CDDP的敏感度,其機(jī)制可能與FANCD2 sh RNA干擾引起B(yǎng)ax/bcl-2、Cyclin D1、HIF-1α的表達(dá)變化有關(guān)。
[Abstract]:Objective to study the effect of FANCD2 sh RNA interference on chemotherapeutic sensitivity of human head and neck squamous cell carcinoma (head and neck squamous cell carcinoma,HNSCC) cell line SIHN-005A and its possible mechanism. Methods Human HNSCC SIHN-005A cells with stable effect of FANCD2 gene silencing were used to detect the silencing effect by purine mycin continuous screening; Western blot, the inhibition rate of cell proliferation by CCK-8 assay and apoptosis by Hoechst assay. The expression of HIF-1 偽 protein was detected by Western blot. Results the expression of FANCD2 protein decreased significantly in the experimental group, and the silencing efficiency of FANCD2 protein reached 60.5%. The inhibition rate of cell proliferation in the experimental group was significantly higher than that in the negative control group and the blank control group, and was in a concentration and time dependent manner. The IC50 value of CDDP in the experimental group was significantly lower than that in the negative control group and the blank control group. The apoptosis rate of the experimental group was increased after FANCD2 gene silencing. After adding CDDP, the expression of Bcl-2HIF-1 偽 protein in Cyclin D1 was significantly down-regulated and Bax protein was significantly increased compared with that before CDDP. Conclusion silencing FANCD2 can increase the sensitivity of SIHN-005A cells to the chemotherapeutic drug CDDP, and the mechanism may be related to the change of Bax/bcl-2,Cyclin D1HIF-1 偽 expression induced by FANCD2 sh RNA interference.
【作者單位】： 西南醫(yī)科大學(xué)附屬醫(yī)院耳鼻咽喉頭頸外科;成都市第一人民醫(yī)院耳鼻咽喉科;
【基金】：四川省教育廳重點(diǎn)項(xiàng)目(12ZA076) 四川省衛(wèi)生廳科研項(xiàng)目(120352) 瀘州醫(yī)學(xué)院附屬醫(yī)院留學(xué)歸國(guó)人員科研項(xiàng)目([2013]60-2) 四川省科技廳-瀘州市科技局-瀘州醫(yī)學(xué)院聯(lián)合科研項(xiàng)目(14JC0182)
【分類號(hào)】：R739.91

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本文編號(hào)：2330888