MicroRNA-27b介导沉默同源盒蛋白HOXB8对人胃癌干细胞样特性的调节作用

doi:10.3969/j.issn.1006-6233.2021.11.02

Abstract
Figure/Table
References
Related Citation (0)

Download: PDF (2312 KB) HTML (1 KB)
Export: BibTeX | EndNote (RIS)

Abstract Objective: To explore the expression levels of miR-27b and homeobox protein HOXB8 in gastric cancer tissues, and the mechanisms by which they regulate human gastric cancer stem cell-like characteristics. Methods: Collected 30 cases of gastric cancer tissues and corresponding normal tissues adjacent to the cancer in our hospital. The differences in the expression of miR-27b and HOXB8 in tissues was detected with real-time fluorescence quantitative PCR. Bioinformatics tool TargetScan7.2 and dual luciferase reporter gene technology were adopted to predict and verify the targeting relationship between miR-27b and HOXB8. The immunomagnetic bead sorting method was used to separate gastric cancer stem cells from the gastric cancer cell line SGC-7901. MiR-27b mimic and negative control (miR-27b NC) were transfected into gastric cancer stem cells by liposome-mediated method. The expression of miR-27b in gastric cancer stem cells after transfection was detected with real-time fluorescence quantitative PCR, Western blot was adopted to detect HOXB8 protein expression in gastric cancer stem cells after transfection, CCK-8 method was used to detect the activity of gastric cancer stem cells, EdU test was applied to detect the proliferation of gastric cancer stem cells, Cell spheroidization experiment and cell clone formation experiment were conducted to detect the self-renewal ability of gastric cancer stem cells. Results: The expression level of miR-27b in gastric cancer tissues was higher than that of normal tissues adjacent to cancer, while HOXB8 was lower than that of normal tissues adjacent to cancer (P<0.01); miR-27b and HOXB8 3'-UTR have base complementation in specific regions. The relative fluorescence of cells co-transfected with miR-27b mimic and WT-HOXB8 3’-UTR recombinant plasmid was lower than that of cells co-transfected with miR-27b NC and WT-HOXB8 3’-UTR recombinant plasmid (P<0.01). After sorting, high-purity CD44+ labeled gastric cancer stem cells were obtained, and miR-27b mimic was successfully transfected into gastric cancer stem cells. Compared with the blank control group, miR-27b expression in gastric cancer stem cells of miR-27b mimic group increased (P<0.01), HOXB8 protein expression decreased (P<0.01), cell viability decreased (P<0.01), EdU-labeled positive cells decreased (P<0.01), the number of cells into spheres is less, the size of spheres becomes smaller, and the number of cell clone formation decreased (P<0.01). Conclusion: The expression of miR-27b is decreased and the expression of HOXB8 is increased in gastric cancer tissues. MiR-27b regulates the proliferation and self-renewal ability of gastric cancer stem-like cells by targeting to silence the expression of HOXB8.

Key words： Gastric cancer stem cells MiR-27b Homeobox protein 8 Stem cell-like characteristics

	Service

	E-mail this article
	Add to my bookshelf
	Add to citation manager
	E-mail Alert
	RSS
	Articles by authors
	YANG Xiangchao
	TIAN Youjing
	CHEN Yu
	et al

Cite this article:

YANG Xiangchao,TIAN Youjing,CHEN Yu, et al. MicroRNA-27b-Mediated Silencing of Homeobox Protein HOXB8 Modulates Human Gastric Cancer Stem Cell-Like Characteristics[J]. HeBei Med, 2021, 27(11): 1766-1773.

URL:

http://www.hbyxzzs.cn/EN/10.3969/j.issn.1006-6233.2021.11.02 OR http://www.hbyxzzs.cn/EN/Y2021/V27/I11/1766

[1] Smyth EC,Nilsson M,Grabsch HI,et al.Gastric cancer[J].LANCET,2020,396 (10251):635~648.
[2] Yasui M,Kunita A,Numakura S,et al.Cancer stem cells in Epstein-Barr virus-associated gastric carcinoma[J].Cancer Sci,2020,111(7):2598~2607.
[3] Dilsiz N.Role of exosomes and exosomal microRNAs in cancer[J].Future Sci OA,2020,6(4):46~52.
[4] Li D,Jie N,Fan Y,et al.Promising therapeutic role of miR-27b in tumor[J].Tumor Biol,2017,39(3):87~108.
[5] Ying Y,Wang Y,Huang X,et al.Oncogenic HOXB8 is driven by MYC-regulated super-enhancer and potentiates colorectal cancer invasiveness via BACH1[J].Oncogene,2020,39(5):1004~1017.
[6] Chu JY,McCormick B,Mazelyte G,et al.HoxB8 neutrophils replicate Fcγ receptor and integrin-induced neutrophil signaling and functions[J].Leukoc Biol,2019,105(1):93~100.
[7] Sohn SH,Kim B,Sul HJ,et al.Foretinib inhibits cancer stemness and gastric cancer cell proliferation by decreasing CD44 and c-MET signaling[J].Onco Targets Ther,2020,13:1027~1035.
[8] Bekaii-Saab T,El-Rayes B.Identifying and targeting cancer stem cells in the treatment of gastric cancer[J].Cancer,2017,123(8):1303~1312.
[9] Kawaguchi M,Dashzeveg N,Cao Y,et al.Extracellular domains I and II of cell-surface glycoprotein CD44 mediate its trans-homophilic dimerization and tumor cluster aggregation[J].Biol Chem,2020,295(9):2640~2649.
[10] Zhang J,Hua X,Qi N,et al.MiR-27b suppresses epithelial-mesenchymal transition and chemoresistance in lung cancer by targeting Snail1[J].Life Sci,2020,254:117238.
[11] Feng Q,Wu X,Li F,et al.miR-27b inhibits gastric cancer metastasis by targeting NR2F2[J].Protein Cell,2017,8 (2):114~122.
[12] Ying Y,Wang Y,Huang X,et al.Oncogenic HOXB8 is driven by MYC-regulated superenhancer and potentiates colorectal cancer invasiveness via BACH1[J].Oncogene,2020,39(5):1004~1177.
[13] Guo J,Zhang T,Dou D.Knockdown of HOXB8 inhibits tumor growth and metastasis by the inactivation of Wnt/β-catenin signaling pathway in osteosarcoma[J].Eur Pharmacol,2019,854:22~27.
[14] Liu YJ,Zhou HG,Chen LH,Qu DC,Wang CJ,Xia ZY,Zheng JH.MiR-32-5p regulates the proliferation and metastasis of cervical cancer cells by targeting HOXB8[J].Eur Rev Med Pharmacol Sci,2019,23(1):87~95.
[15] Ding WJ,Zhou M,Chen MM,et al.HOXB8 promotes tumor metastasis and the epithelial~mesenchymal transition via ZEB2 targets in gastric cancer[J].Cancer Res Clin Oncol,2017,143(3):385~397.