摘要
目的:探讨长链非编码RNA-linc00346调控膀胱癌细胞增殖的分子机制。方法:通过实时荧光定量聚合酶链反应(RT-PCR)检测膀胱尿路上皮癌组织中linc00346 mRNA和PTEN mRNA的表达情况;沉默膀胱癌T24细胞系内linc00346的表达,通过RT-PCR检测T24细胞内的linc00346 mRNA和PTEN mRNA表达变化情况,通过蛋白质印迹(Western blot)法检测T24细胞内PTEN蛋白和PI3K/Akt信号通路中Akt、p-Akt蛋白的表达情况;CCK-8实验检测各组T24细胞的增殖能力。结果:与膀胱正常组织比较,膀胱尿路上皮癌组织中linc00346 mRNA表达上调,而PTEN mRNA的表达水平降低(均P<0.05)。沉默linc00346表达后,T24细胞内linc00346 mRNA的表达水平显著下降,PTEN mRNA的表达水平显著上调,PTEN蛋白水平升高,p-Akt蛋白水平降低(均P<0.05);给予PTEN特异性抑制剂SF1670处理后,PTEN蛋白水平显著下调,p-Akt蛋白水平明显增加(均P<0.05)。沉默linc00346表达后,T24细胞的增殖能力明显受到抑制,给予SF1670处理后细胞的增殖能力显著增强(均P<0.05)。结论:linc00346可以增强膀胱尿路上皮癌细胞的增殖能力,其作用机制可能是通过抑制PTEN表达,激活PI3K/Akt信号通路实现的。
Objective: To study the molecular mechanism of linc00346 in promoting proliferation of bladder cancer cells. Methods: Real-time fluorescence quantitative PCR(RT-PCR) was used to detect the expression of linc00346 and PTEN in urothelial carcinoma tissues of bladder. The expression level of linc00346 and PTEN in bladder cancer cell lines T24 was detected by RT-PCR after knockdown linc00346. The expression of PTEN protein and Akt, p-Akt protein in PI3 K/Akt signaling pathway was measured by Western blot;The cell proliferation ability was determined by CCK-8 method. Results: The expression of linc00346 was up-regulated, while the PTEN expression level was down-regulated in urothelial carcinoma tissues(P<0.05). After knockdown linc00346 in T24 cells, the expression level of linc00346 decreased, the level of PTEN mRNA increased(P<0.05), the expression of PTEN protein increased, and the expression of p-Akt reduced(P<0.05). When PTEN inhibitor SF1670 was given at the same time, the expression of PTEN protein was down-regulated, and that of p-Akt was up-regulated(P<0.05). The proliferation ability of T24 cells was inhibited obviously after silencing linc00346.Furthermore, SF1670 made the cell proliferation ability increased(P<0.05). Conclusion: Linc00346 can increase the proliferation ability of bladder urothelial carcinoma cells by inhibiting PTEN, which activates PI3 K/Akt signaling pathway.
作者
叶挺宇
黄航
潘悦
蔡冰
YE Tingyu;HUANG Hang;PAN Yue;CAI Bing(Department of Urology,the First Affiliated Hospital of Wenzhou Medical University,Wenzhou 325015,China)
出处
《温州医科大学学报》
2021年第12期995-999,共5页
Journal of Wenzhou Medical University
基金
温州市基础性科研项目(Y20190567)。
