RNAi NRP-1基因对骨肉瘤细胞凋亡、ROS含量及STAT3信号通路的影响-东南大学学报医学版

RNAi NRP-1基因对骨肉瘤细胞凋亡、ROS含量及STAT3信号通路的影响

单位：1. 海南省第三人民医院骨科, 海南三亚 572000;
2. 北京大学人民医院骨肿瘤科, 北京 100044

分类号：R738.6

出版年·卷·期（页码）：2019·38·第一期（1-5）

摘要：

目的：探讨RNAi神经纤毛蛋白1（NRP1）基因对骨肉瘤细胞凋亡、ROS含量及STAT3信号通路的影响。方法：分别将合成的NRP-1的siRNA（NRP-1-siRNA组）及无干扰作用的siRNA（NC组）转染人骨肉瘤MG-63细胞，并设置空白对照组，Western blotting检测转染48 h的细胞中NRP-1和磷酸化的STAT3（p-STAT3）及STAT3信号通路靶基因Cyclin D1和Bcl-2的蛋白表达；CCK8法分别于转染的24、48 h和72 h检测细胞活力；流式细胞术分别检测细胞凋亡率及ROS含量。结果：与空白对照组比较，NRP-1-siRNA组NRP-1、Cyclin D1、Bcl-2和p-STAT3的蛋白表达均显著降低，转染24、48、72 h的细胞活力均显著降低，细胞凋亡率显著升高，ROS含量显著升高（P<0.05）。结论：RNAi抑制NRP-1基因表达可降低骨肉瘤细胞活力，诱导细胞凋亡，机制可能与提高细胞内ROS含量和下调STAT3信号通路有关。

Objective:To investigate the effect of NRP-1 gene by RNA interference on apoptosis, ROS content and STAT3 signaling pathway in osteosarcoma cells. Methods:siRNA of synthetic NRP-1 (NRP-1-siRNA group) and siRNA without interference (NC group) were transfected to human osteosarcoma MG-63 cells, respectively.Western blotting was used to detect the expression of NRP-1, STAT3, phosphorylated STAT3(p-STAT3) and STAT3 signaling pathway target genes Cyclin D1 and Bcl-2 protein cells transfected for 48 h;CCK8 assay was used to detect cell viability cells transfected 24 h, 48 h and 72 h, respectively; cell apoptosis rate and ROS content were detected by flow cytometry. Results:Compared with the blank control group, the protein expression of NRP-1, Cyclin D1, Bcl-2 and p-STAT3 in NRP-1-siRNA group decreased significantly, and the cell viability decreased significantly at three time points, the apoptosis rate and ROS content increased significantly (P<0.05). Conclusion:Inhibition of NRP-1 gene expression by RNA interference can reduce osteosarcoma cell viability, induce cell apoptosis, the mechanism may be related to increase ROS content and downregulate STAT3 signaling pathway.

参考文献：

[1] SHANG G,MI Y,MEI Y,et al.MicroRNA-192 inhibits the proliferation,migration and invasion of osteosarcoma cells and promotes apoptosis by targeting matrix metalloproteinase-11[J].Oncol Lett,2018,15(5):7265-7272.
[2] CHU W,SONG X,YANG X,et al.Neuropilin-1 promotes epithelial-to-mesenchymal transition by stimulating nuclear factor-kappa B and is associated with poor prognosis in human oral squamous cell carcinoma[J].PLoS One,2014,9(7):e101931.
[3] KERROS C,TRIPATHI S C,ZHA D,et al.Neuropilin-1 mediates neutrophil elastase uptake and cross-presentation in breast cancer cells[J].J Biol Chem,2017,292(24):10295-10305.
[4] KWIATKOWSKI S C,GUERRERO P A,HIROTA S,et al.Neuropilin-1 modulates TGFβ signaling to drive glioblastoma growth and recurrence after anti-angiogenic therapy[J].PLoS One,2017,12(9):e0185065.
[5] SUN J,WANG L,LOU W,et al.Construction and identification of small hairpin RNA plasmids targeting neuropilin-1 gene and their inhibitory effect on human nasopharyngeal carcinoma CNE-2Z cell proliferation in vitro and in vivo[J].Eur Arch Otorhinolaryngol,2016,273(9):2541-2547.
[6] YUE B,MA J F,YAO G,et al.Knockdown of neuropilin-1 suppresses invasion,angiogenesis,and increases the chemosensitivity to doxorubicin in osteosarcoma cells-an in vitro study[J].Eur Rev Med Pharmacol Sci,2014,18(12):1735-1741.
[7] ZHU H,CAI H,TANG M,et al.Neuropilin-1 is overexpressed in osteosarcoma and contributes to tumor progression and poor prognosis[J].Clin Transl Oncol,2014,16(8):732-738.
[8] LIU Y,BI T,DAI W,et al.Lupeol induces apoptosis and cell cycle arrest of human osteosarcoma cells through PI3K/AKT/mTOR pathway[J].Technol Cancer Res Treat,2016,15(6):NP16-NP24.
[9] XU B,XIA H,CAO J,et al.MicroRNA-21 inhibits the apoptosis of osteosarcoma cell line SAOS-2 via targeting caspase 8[J].Oncol Res,2017,25(7):1161-1168.
[10] AKASHI Y,ODA T,OHARA Y,et al.Anticancer effects of gemcitabine are enhanced by co-administered iRGD peptide in murine pancreatic cancer models that overexpressed neuropilin-1[J].Br J Cancer,2014,110(6):1481-1487.
[11] KUMAR A,HUO S,ZHANG X,et al.Neuropilin-1-targeted gold nanoparticles enhance therapeutic efficacy of platinum (IV) drug for prostate cancer treatment[J].ACS Nano,2014,8(5):4205-4220.
[12] SHI F,SHANG L,PAN B Q,et al.Calreticulin promotes migration and invasion of esophageal cancer cells by upregulating neuropilin-1 expression via STAT5A[J].Clin Cancer Res,2014,20(23):6153-6162.
[13] MARIC G,ANNIS M G,DONG Z,et al.GPNMB cooperates with neuropilin-1 to promote mammary tumor growth and engages integrin α5β1 for efficient breast cancer metastasis[J].Oncogene,2015,34(43):5494-5504.
[14] DU L,FAN Q,TU B,et al.Establishment and characterization of a new highly metastatic human osteosarcoma cell line derived from Saos2[J].Int J Clin Exp Pathol,2014,7(6):2871-2882.
[15] 李天客,陈中,包阳,等.RNA干扰SBF2对人口腔癌细胞株SCC15增殖,凋亡及侵袭的影响[J].中国老年学杂志,2016,36(21):5260-5262.
[16] BOREL F,KAY M A,MUELLER C.Recombinant AAV as a platform for translating the therapeutic potential of RNA interference[J].Mol Ther,2014,22(4):692-701.
[17] 张志强,高体云,李暐,等.雷公藤红素通过活性氧诱导结肠癌SW620细胞凋亡[J].中国医院药学杂志,2015,35(17):1558-1563.
[18] SHI Y,NIKULENKOV F,ZAWACKA-PANKAU J,et al.ROS-dependent activation of JNK converts p53 into an efficient inhibitor of oncogenes leading to robust apoptosis[J].Cell Death Dis,2014,21(4):612-623.
[19] 徐长华,邹明花,张献全.3,3'二吲哚甲烷阻断STAT3通路影响卵巢癌细胞增殖,凋亡的实验研究[J].第三军医大学学报,2014,36(16):1674-1678.
[20] XIONG M,WANG L,YU H L,et al.Ginkgetin exerts growth inhibitory and apoptotic effects on osteosarcoma cells through inhibition of STAT3 and activation of caspase-3/9[J].Oncol Rep,2016,35(2):1034-1040.
[21] LI W,SAUD S M,YOUNG M R,et al.Cryptotanshinone,a Stat3 inhibitor,suppresses colorectal cancer proliferation and growth in vitro[J].Mol Cell Biochem,2015,406(1-2):63-73.
[22] QIU R,MA G,ZHENG C,et al.Antineoplastic effect of calycosin on osteosarcoma through inducing apoptosis showing in vitro and in vivo investigations[J].Exp Mol Pathol,2014,97(1):17-22.

服务与反馈：

【文章下载】【发表评论】【查看评论】【加入收藏】

提示：您还未登录，请登录！点此登录