LINC00662通过调控miR-215-3p/EIF4H轴影响骨肉瘤细胞的增殖和凋亡-现代医学

LINC00662通过调控miR-215-3p/EIF4H轴影响骨肉瘤细胞的增殖和凋亡

单位：华中科技大学协和东西湖医院/武汉市东西湖区人民医院骨科, 湖北武汉 430040

分类号：R738.1

出版年·卷·期（页码）：2021·49·第十期（1211-1218）

摘要：

目的：探讨长基因间非蛋白质编码RNA 662（LINC00662）对骨肉瘤细胞的增殖和凋亡的影响及可能机制。方法：实时荧光定量聚合酶链反应（RT-qPCR）和Western blotting检测人成骨细胞株hFOB 1.19和骨肉瘤细胞中LINC00662、miR-215-3p和真核生物翻译起始因子4H （EIF4H）表达水平。将LINC00662小干扰RNA （si-LINC00662）、miR-215-3p模拟物、EIF4H小干扰RNA （si-EIF4H）分别转染Saos2细胞，细胞计数试剂盒（CCK-8）检测细胞活力，流式细胞术检测细胞凋亡。双荧光素酶报告实验、RT-qPCR、Western blotting确定LINC00662与miR-215-3p、miR-215-3p与EIF4H之间的靶向关系。结果：与hFOB 1.19比较，骨肉瘤细胞中LINC00662和EIF4H表达显著升高，miR-215-3p表达显著降低（P<0.05）。抑制LINC00662表达或过表达miR-215-3p或抑制EIF4H表达后，Saos2细胞存活率均显著降低，凋亡率均显著升高（P<0.05）。LINC00662靶向负性调控miR-215-3p表达，miR-215-3p靶向负性调控EIF4H表达（P<0.05）。抑制miR-215-3p表达可逆转抑制miR-215-3p对Saos2细胞存活和凋亡的影响（P<0.05）。过表达EIF4H可逆转抑制miR-215-3p对Saos2细胞存活和凋亡的影响（P<0.05）。结论：干扰LINC00662表达可抑制骨肉瘤细胞增殖，促进细胞凋亡，其机制与靶向调控miR-215-3p/EIF4H轴有关。

Objective: To investigate the effect of long intergene non-protein coding RNA 662(LINC00662) on the proliferation and apoptosis of osteosarcoma cells, and to explore its possible mechanism further. Methods: Real-time fluorescence quantitative PCR(RT-qPCR) and Western blotting were used to detect the expression level of LINC00662, miR-215-3p and Eukaryotic translation initiation factor 4H(EIF4H) in human osteoblast cell line hFOB 1.19 and osteosarcoma cells. LINC00662 small interfering RNA(si-LINC00662), miR-215-3p mimic, EIF4H small interfering RNA(si-EIF4H) were transfected into Saos2 cells, respectively. Cell counting kit(CCK-8) detected cell viability andflow cytometry detected cell apoptosis. The dual luciferase report experiment, RT-qPCR and Western blotting were applied to confirm the targeting relationship between LINC00662 and miR-215-3p,miR-215-3p and EIF4H. Results: Compared with hFOB 1.19, the expression of LINC00662 and EIF4H in osteosarcoma cells were significantly increased, while the expression of miR-215-3p was significantly reduced(P<0.05). After inhibiting the expression of LINC00662 or overexpressing miR-215-3p or inhibiting the expression of EIF4H, the survival rate of Saos2 cells was significantly reduced, and the apoptosis rate was significantly increased(P<0.05). LINC00662 targets and negatively regulates miR-215-3p expression, and miR-215-3p targets and negatively regulates EIF4H expression(P<0.05). Inhibiting the expression of miR-215-3p could reverse the effect of inhibiting miR-215-3p on the survival and apoptosis of Saos2 cells(P<0.05). Overexpression of EIF4H could reverse the effect of inhibiting miR-215-3p on the survival and apoptosis of Saos2 cells(P<0.05). Conclusion: Interfering with the expression of LINC00662 could inhibit osteosarcoma cells proliferation and promote cell apoptosis, and its mechanism was related to the targeted regulation of miR-215-3p/EIF4H axis.

参考文献：

[1] BERNER K, JOHANNESEN T B, BERNER A, et al. Time-trends on incidence and survival in a nationwide and unselected cohort of patients with skeletal osteosarcoma[J]. Acta Oncol, 2015, 54(1):25-33.
[2] FRIEBELE J C, PECK J, PAN X, et al. Osteosarcoma:a meta-analysis and review of the literature[J]. Am J Orthop, 2015, 44(12):547-553.
[3] MA J, YANG Y, HUO D, et al. LincRNA-RoR/miR-145 promote invasion and metastasis in triple-negative breast cancer via targeting MUC1[J]. Biochem Biophys Res Commun, 2018, 500(3):614-620.
[4] 兰飞, 赵鹏, 刘冬, 等.LncRNA DANCR调控非小细胞肺癌细胞的凋亡和上皮间质转化[J]. 现代医学, 2020, 48(1):81-88.
[5] XU D, CHEN Y, YUAN C, et al. Long non-coding RNA LINC00662 promotes proliferation and migration in oral squamous cell carcinoma[J]. Onco Targets Ther, 2019, 12(1):647-656.
[6] LIU Y, GAO X, TIAN X.High expression of long intergenic non-coding RNA LINC00662 contributes to malignant growth of acute myeloid leukemia cells by upregulating ROCK1 via sponging microRNA-340-5p[J]. Eur J Pharmacol, 2019, 859(1):172535-172544.
[7] LIU C Q, CHEN Y, XIE B F, et al. MicroRNA-215-3p suppresses the growth and metastasis of cervical cancer cell via targeting SOX9[J]. Eur Rev Med Pharmacol Sci, 2019, 23(13):5628-5639.
[8] BAI Y, LU C, ZHANG G, et al. Overexpression of miR-519d in lung adenocarcinoma inhibits cell proliferation and invasion via the association of eIF4H[J]. Tumour Biol, 2017, 39(3):1010428317694566.
[9] KONG G, QI X J, WANG J F.Effect of lncRNA LET on proliferation and invasion of osteosarcoma cells[J]. Eur Rev Med Pharmacol Sci, 2018, 22(6):1609-1614.
[10] 赵娟娟, 韦四喜, 严芝强, 等.胃癌长链非编码RNA SNHG16异常表达及其临床病理意义[J]. 东南大学学报(医学版), 2017, 36(4):551-554.
[11] LI N, ZHANG L Y, QIAO Y H, et al. Long noncoding RNA LINC00662 functions as miRNA sponge to promote the prostate cancer tumorigenesis through targeting miR-34a[J]. Eur Rev Med Pharmacol Sci, 2019, 23(9):3688-3698.
[12] GONG W, SU Y, LIU Y, et al. Long non-coding RNA Linc00662 promotes cell invasion and contributes to cancer stem cell-like phenotypes in lung cancer cells[J]. J Biochem, 2018, 164(6):461-469.
[13] TIAN X, WU Y, YANG Y, et al. Long noncoding RNA LINC00662 promotes M2 macrophage polarization and hepatocellular carcinoma progression via activating Wnt/β-catenin signaling[J]. Mol Oncol, 2020, 14(2):462-483.
[14] CHENG B, RONG A, ZHOU Q, et al. LncRNA LINC00662 promotes colon cancer tumor growth and metastasis by competitively binding with miR-340-5p to regulate CLDN8/IL22 co-expression and activating ERK signaling pathway[J]. J Exp Clin Cancer Res, 2020, 39(1):5-15.
[15] LIU Z, YAO Y, HUANG S, et al. LINC00662 promotes gastric cancer cell growth by modulating the Hippo-YAP1 pathway[J]. Biochem Biophys Res Commun, 2018, 505(3):843-849.
[16] QI X, ZHANG DH, WU N, et al. ceRNA in cancer:possible functions and clinical implications[J]. J Med Genet, 2015, 52(10):710-718.
[17] WANG H, YU M, HU W, et al. Linc00662 promotes tumorigenesis and progression by regulating miR-497-5p/AVL9 axis in colorectal cancer[J]. Front Genet, 2020, 10(1):1385-1395.
[18] ZHANG F, WANG X S, TANG B, et al. Long non-coding RNA FTX promotes gastric cancer progression by targeting miR-215[J]. Eur Rev Med Pharmacol Sci, 2020, 24(6):3037-3048.
[19] LI X W, QIU S J, ZHANG X.Overexpression of miR-215-3p sensitizes colorectal cancer to 5-fluorouracil induced apoptosis through regulating CXCR1[J]. Eur Rev Med Pharmacol Sci, 2018, 22(21):7240-7250.
[20] VAYSSE C, PHILIPPE C, MARTINEAU Y, et al. Key contribution of eIF4H-mediated translational control in tumor promotion[J]. Oncotarget, 2015, 6(37):39924-39940.

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