本文選題：miR- + NEK��； 參考：《廣東醫(yī)學(xué)》2017年07期

【摘要】：目的研究miR-195調(diào)控中心體相關(guān)激酶2(NEK2)對前列腺癌細胞增殖的影響。方法 Western blot檢測在穩(wěn)定表達和抑制表達miR-195的LNCaP細胞株中NEK2蛋白的表達水平;構(gòu)建克隆編碼miR-195的序列片段慢病毒載體和表達NEK2的質(zhì)粒載體,對轉(zhuǎn)染空載體和NEK2,以及同時轉(zhuǎn)染了miR-195模擬物和NEK2、miR-195模擬物和空載體的LNCaP細胞株進行CCK8細胞增殖實驗;采用shRNA方法構(gòu)建干擾NEK2基因表達質(zhì)粒,NEK2抑制成功和miR-195過表達的LNCaP細胞進行細胞增殖實驗,測定si-NC、si-NEK2、si-195的LNCaP細胞的生長速度。結(jié)果在過表達miR-195的LNCaP細胞株中NEK2蛋白的表達水平比陰性對照miR-NC的LNCaP細胞株中蛋白表達水平明顯降低(P0.01);相反,在miR-195被抑制表達的LNCaP細胞株中NEK2蛋白的表達水平明顯增加(P0.01)。細胞增殖實驗顯示轉(zhuǎn)染NEK2基因的LNCaP細胞在72、96h生長速度顯著快于轉(zhuǎn)染空載體的細胞(P0.01);轉(zhuǎn)染miR-195模擬物和NEK2的LNCaP細胞在72、96h生長速度顯著慢于單純轉(zhuǎn)染NEK2的LNCaP細胞(P0.01);si-NEK2的LNCaP細胞在72、96h生長速度明顯慢于siNC,且與miR-195過表達的LNCaP細胞相似。結(jié)論 miR-195負性調(diào)控相關(guān)下游基因NEK2的表達,抑制前列腺癌細胞的增殖,初步驗證了miRNA-195-NEK2信號軸在前列腺癌中的作用。
[Abstract]:Objective to study the effect of miR-195 on the proliferation of prostate cancer cells. Methods Western blot was used to detect the expression level of NEK2 protein in the LNCaP cell line expressing miR-195 stably and to inhibit the expression of NEK2 protein. The sequence fragment lentivirus vector encoding miR-195 and the plasmid vector expressing NEK2 were constructed. The proliferation of LNCaP cell lines transfected with empty vector and NEK2, as well as miR-195 mimics and NEK2 miR-195 mimics and empty vectors were studied. ShRNA method was used to construct LNCaP cells which interfered with the expression of NEK2 gene and miR-195 overexpression. The growth rate of LNCaP cells of si-NCCnsi-NEK2 si-195 was measured. Results the expression level of NEK2 protein in LNCaP cell line with overexpression of miR-195 was significantly lower than that in LNCaP cell line with negative control miR-NC, but the expression level of NEK2 protein was significantly increased in LNCaP cell line with inhibited miR-195 expression. Cell proliferation test showed that the growth rate of LNCaP cells transfected with NEK2 gene was significantly faster than that of empty vector transfected LNCaP cells at 72 ~ 96 h, and the growth rate of LNCaP cells transfected with miR-195 mimics and NEK2 at 72 ~ 96 h was significantly slower than that of LNCaP cells transfected with NEK2 alone at 72 ~ 96 h. The growth rate of LNCaP cells transfected with miR-195 mimics and NEK2 was significantly slower than that of LNCaP cells transfected with NEK2 alone. The growth rate of SNC cells was significantly slower than that of siNCcells at 72 ~ 96 h, and was similar to that of LNCaP cells overexpressed by miR-195. Conclusion miR-195 negatively regulates the expression of downstream gene NEK2 and inhibits the proliferation of prostate cancer cells, which preliminarily verifies the role of miRNA-195-NEK2 signaling axis in prostate cancer.
【作者單位】： 廣州醫(yī)科大學(xué)附屬第五醫(yī)院泌尿外科;廣州醫(yī)科大學(xué)附屬廣州市第一人民醫(yī)院泌尿外科;廣州醫(yī)科大學(xué)附屬廣州市第一人民醫(yī)院麻醉科;
【基金】：廣東省自然科學(xué)基金資助項目(編號:2014A030310501) 廣州市醫(yī)藥衛(wèi)生科技項目(編號:20141A011006)
【分類號】：R737.25

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本文編號：1950671