miR-26b对口腔鳞癌细胞SCC15肿瘤干性的抑制作用
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摘要
目的分析miR-26b对人口腔鳞癌细胞SCC15肿瘤干性的影响及潜在机制。方法比较5例口腔癌患者癌组织和癌旁组织miR-26b表达差异;以miR-26b模拟物及抑制物瞬染SCC15细胞,Sphere实验检测癌细胞体外成球能力、real-time PCR检测干性基因(Nanog/OCT4)表达,Western Blot检测上皮间质转化标志物E-钙粘蛋白水平。结果 5例患者癌组织miR-26b水平均低于癌旁组织(P <0.05)。过表达miR-26b使SCC15细胞成球率下降(P <0.05),Nanog及OCT4表达下调(P <0.05),E-钙粘蛋白水平升高(P <0.05);反之抑表达miR-26b后,SCC15成球率上升(P <0.05),Nanog及OCT4表达上升(P <0.05),E-钙粘蛋白水平下降(P <0.05)。结论 miR-26b能降低SCC15细胞的肿瘤干性,其机制可能与miR-26b抑制细胞的上皮间质转化有关。
Objective To investigate the effect of miR-26 b on SCC15 cell′s tumor stemness and the possible mechanism of its effect. Methods Real-time PCR was used to detect differences of miR-26 b expression in SCC15 cells of cancerous tissues and adjacent tissues of 5 oral carcinoma patients. Real-time PCR was also used to detect the stem gene(Nanog/OCT4)of SCC15 cells. The miR-26 b mimics and miR-26 b inhibitors were respectively transected into SCC15 cells. Sphere experiment was used to detect in-vitro sphere-forming ability of SCC15 cells.E-cadherin protein level was measured by Western Blot. Results Compared with adjacent tissues,miR-26 b expression of cancerous tissues is significantly lower(P < 0.05). In SCC15 cells with miR-26 b overexpression,the sphere-forming ability decreased(P < 0.05);Nanog and OCT4 expression were reduced(P < 0.05);E-cadherin level was increased(P < 0.05). In contrast,in SCC15 cells with miR-26 b inhibition,the sphere-forming ability increased(P < 0.05);Nanog and OCT4 expression were enhanced(P < 0.05);E-cadherin level was increased(P < 0.05). Conclusion miR-26 b suppresses the tumor cell stemness of SCC15 cells. The mechanism may be related to inhibition of miR-26 b on epithelial-mesenchymal transformation in tumor cells.
Objective To investigate the effect of miR-26 b on SCC15 cell′s tumor stemness and the possible mechanism of its effect. Methods Real-time PCR was used to detect differences of miR-26 b expression in SCC15 cells of cancerous tissues and adjacent tissues of 5 oral carcinoma patients. Real-time PCR was also used to detect the stem gene(Nanog/OCT4)of SCC15 cells. The miR-26 b mimics and miR-26 b inhibitors were respectively transected into SCC15 cells. Sphere experiment was used to detect in-vitro sphere-forming ability of SCC15 cells.E-cadherin protein level was measured by Western Blot. Results Compared with adjacent tissues,miR-26 b expression of cancerous tissues is significantly lower(P < 0.05). In SCC15 cells with miR-26 b overexpression,the sphere-forming ability decreased(P < 0.05);Nanog and OCT4 expression were reduced(P < 0.05);E-cadherin level was increased(P < 0.05). In contrast,in SCC15 cells with miR-26 b inhibition,the sphere-forming ability increased(P < 0.05);Nanog and OCT4 expression were enhanced(P < 0.05);E-cadherin level was increased(P < 0.05). Conclusion miR-26 b suppresses the tumor cell stemness of SCC15 cells. The mechanism may be related to inhibition of miR-26 b on epithelial-mesenchymal transformation in tumor cells.
引文
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[3] MITRA A,MISHRA L,LI S. EMT,CTCs and CSCs in tumor relapse and drug-resistance[J]. Oncotarget,2015,6(13):10697-10711.
[4] FAN D,LIN X,ZHANG F,et al. MicroRNA-26b promotes colorectal cancer metastasis by down-regulating PTEN and WNT5A[J]. Cancer Sci,2017,109(2):354-362.
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[7] MAYAKANNAN M,KUHA D M R A,GANESAN A,et al.Oral squamous cell carcinoma:microRNA expression profiling and integrative analyses for elucidation of tumourigenesis mechanism[J]. Mol Cancer,2016,15(1):28.
[8] BAGHAEI F,ABDOLLAHI A,MOHAMMADPOUR H,et al.PTEN and miR-26b:Promising prognostic biomarkers in initiation and progression of oral squamous cell carcinoma[J]. J Oral Pathol Med,2019,48(1):31-35.
[9] LI J,LIANG Y,LV H,et al. miR-26a and miR-26b inhibit esophageal squamous cancer cell proliferation through suppression of c-MYC pathway[J]. Gene,2017,625:1-9.
[10] NAIK P P,MUKHOPADHYAY S,PANDA P K,et al. Autophagy regulates cisplatin-induced stemness and chemoresistance via the upregulation of CD44,ABCB1 and ADAM17 in oral squamous cell carcinoma[J]. Cell Prolif,2018,51(1):e12411.
[11] ZIMMERMAN A L,WU S. MicroRNAs,cancer and cancer stem cells[J]. Cancer Lett,2011,300(1):10-19.
[12] TROMPETER H I,DREESEN J,HERMANN E,et al. MicroRNAs miR-26a,miR-26b,and miR-29b accelerate osteogenic differentiation of unrestricted somatic stem cells from human cord blood[J]. BMC Genomics,2013,14(1):111.
[13] LIN J X,ZHANG L,HUANG H,et al. MiR-26b/KPNA2 axis inhibits epithelial ovarian carcinoma proliferation and metastasis through downregulating OCT4[J]. Oncotarget,2015,6(27):23793-23806.
[14] PARK Y M,LEE S Y,PARK S W,et al. Role of cancer stem cell in radioresistant head and neck cancer[J]. Auris Nasus Larynx,2016,43(5):556-561.
[15] YIN X,ZHANG B H,ZHENG S S,et al. Coexpression of gene Oct4 and Nanog initiates stem cell characteristics in hepatocellular carcinoma and promotes epithelial-mesenchymal transition through activation of Stat3/Snail signaling[J]. J Hematol Oncol,2015,8(1):1-13.
[16] FANTOZZI A,GRUBER D C,PISARSKY L,et al. VEGF-mediated angiogenesis links EMT-induced cancer stemness to tumor initiation[J]. Cancer Res,2014,74(5):1566-1575.
[17] KYOUNG-OK H,KYU-YOUNG O,WUI-JUNG S,et al. Tumor budding is associated with poor prognosis of oral squamous cell carcinoma and histologically represents an epithelial–mesenchymal transition process[J]. Hum Pathol,2018,80:123-129.
[18] OZAKI-HONDA Y,SEKI S,FUJIWARA M,et al. Prognostic prediction of oral squamous cell carcinoma by E-cadherin and Ncadherin expression in overall cells in tumor nests or tumor cells at the invasive front[J]. Cancer Microenviron,2017,10(1-3):87-94.