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LncRNAMCM3AP-AS1/miR-335对骨肉瘤细胞增殖及凋亡的影响
作者:刘元豪  邢忠  吴玮杰  许环顺 
单位:海口市第三人民医院 骨科, 海南 海口 571100
关键词:长链非编码 RNA MCM3AP反义RNA 1 miR-335 骨肉瘤 增殖 凋亡 
分类号:R738
出版年·卷·期(页码):2022·50·第三期(325-329)
摘要:

目的:探讨长链非编码 RNA MCM3AP 反义 RNA 1(LncRNA MCM3AP-AS1)对骨肉瘤细胞增殖、凋亡的影响及其对miR-335的调控作用。方法:采用实时荧光定量聚合酶链反应(qRT-PCR)法检测骨肉瘤组织、瘤旁组织中MCM3AP-AS1、miR-335的表达量;体外培养人骨肉瘤细胞U2OS,分别将si-NC、si-MCM3AP-AS1、si-MCM3AP-AS1与NC-inhibitor、si-MCM3AP-AS1与miR-335 inhibitor转染至U2OS细胞;采用qRT-PCR法检测U2OS细胞中MCM3AP-AS1、miR-335的表达量;采用MTT实验与平板克隆形成实验检测细胞增殖能力;采用流式细胞术检测细胞凋亡率;双荧光素酶报告实验检测MCM3AP-AS1、miR-335的靶向关系;蛋白质印迹法检测B淋巴细胞瘤-2相关蛋白(Bax)、B淋巴细胞瘤-2(Bcl-2)蛋白表达量。结果:与瘤旁组织比较,骨肉瘤组织中MCM3AP-AS1的表达水平升高(P<0.05),miR-335的表达水平降低(P<0.05);转染si-MCM3AP-AS1可明显降低光密度值及Bcl-2蛋白水平、减少克隆形成数、提高凋亡率及Bax蛋白水平(P<0.05);双荧光素酶报告实验证实MCM3AP-AS1可靶向结合miR-335;转染miR-335 inhibitor可明显逆转干扰MCM3AP-AS1对U2OS细胞增殖及凋亡的作用。结论:MCM3AP-AS1可通过负向调控miR-335的表达调节骨肉瘤细胞增殖及凋亡。

Objective: To explore the effect of long non-coding RNA MCM3AP antisense RNA1(LncRNA MCM3AP-AS1) on the proliferation and apoptosis of osteosarcoma cells and its regulatory effect on miR-335. Methods: The qRT-PCR method was used to detect the expression of MCM3AP-AS1 and miR-335 in osteosarcoma tissue and adjacent tissues. Human osteosarcoma cells U2OS were cultured in vitro, and si-NC, si-MCM3AP-AS1, si-MCM3AP-AS1 and NC-inhibitor, si-MCM3AP-AS1 and miR-335 inhibitor were respectively transfected into U2OS cells. The qRT-PCR method was used to detect the expression of MCM3AP-AS1 and miR-335 in U2OS cells. MTT experiment and plate clone formation experiment were used to detect cell proliferation ability. Flow cytometry was used to detect the apoptosis rate. The dual luciferase reporter experiment was used to detect the targeting relationship of MCM3AP-AS1 and miR-335. Western blot method was used to detect the protein expression of Bax and Bcl-2. Results: Compared with adjacent tissues, the expression level of MCM3AP-AS1 in osteosarcoma tissue was significantly increased(P<0.05), and the expression level of miR-335 was significantly decreased(P<0.05). Transfection of si-MCM3AP-AS1 could significantly reduce the OD value and the protein level of Bcl-2, reduce the number of clones, increase the apoptosis rate and the protein level of Bax(P<0.05). The dual luciferase report experiment confirmed that MCM3AP-AS1 could target miR-335. Transfection of miR-335 inhibitor could significantly reverse the effect of interference MCM3AP-AS1 on the proliferation and apoptosis of U2OS cells. Conclusion: MCM3AP-AS1 can regulate the proliferation and apoptosis of osteosarcoma cells by negatively regulating the expression of miR-335.

参考文献:

[1] 李国湘,方业汉,吴彬.半枝莲碱对人骨肉瘤细胞增殖、侵袭与迁移的抑制作用及其机制[J].东南大学学报(医学版),2021,40(4):456-462.
[2] XIE C,CHEN B,WU B,et al.LncRNA TUG1 promotes cell proliferation and suppresses apoptosis in osteosarcoma by regulating miR-212-3p/FOXA1 axis[J].Biomed Pharmacother,2018,97(1):1645-1653.
[3] LUO X F,WU X J,WEI X,et al.LncRNA ADPGK-AS1 regulated cell proliferation,invasion,migration and apoptosis via targeting miR-542-3p in osteosarcoma[J].Eur Rev Med Pharmacol Sci,2019,23(20):8751-8760.
[4] HUANG Q,YANG J,HE X,et al.LncRNA BDNF-AS is associated with the malignant status and regulates cell proliferation and apoptosis in osteosarcoma[J].Biosci Rep,2018,38(6):1-10.
[5] WANG Y,YANG L,CHEN T,et al.A novel lncRNA MCM3AP-AS1 promotes the growth of hepatocellular carcinoma by targeting miR-194-5p/FOXA1 axis[J].Mol Cancer,2019,18(1):28-38.
[6] LIU J,BIAN T,FENG J,et al.miR-335 inhibited cell proliferation of lung cancer cells by target Tra2β[J].Cancer Sci,2018,109(2):289-296.
[7] YU X,PANG L,YANG T,et al.lncRNA LINC01296 regulates the proliferation,metastasis and cell cycle of osteosarcoma through cyclin D1[J].Oncol Rep,2018,40(5):2507-2514.
[8] JIA D,NIU Y,LI D,et al.lncRNA C2dat1 promotes cell proliferation,migration,and invasion by targeting miR-34a-5p in osteosarcoma cells[J].Oncol Res,2018,26(5):753-764.
[9] YE F,TIAN L,ZHOU Q,et al.LncRNA FER1L4 induces apoptosis and suppresses EMT and the activation of PI3K/AKT pathway in osteosarcoma cells via inhibiting miR-18a-5p to promote SOCS5[J].Gene,2019,721(1):144093-144103.
[10] YANG W,SHAN Z,ZHOU X,et al.Knockdown of lncRNA GHET1 inhibits osteosarcoma cells proliferation,invasion,migration and EMT in vitro and in vivo[J].Cancer Biomark,2018,23(4):589-601.
[11] LIANG M,JIA J,CHEN L,et al.LncRNA MCM3AP-AS1 promotes proliferation and invasion through regulating miR-211-5p/SPARC axis in papillary thyroid cancer[J].Endocrine,2019,65(2):318-326.
[12] ZHANG H,LUO C,ZHANG G.LncRNA MCM3AP-AS1 regulates epidermal growth factor receptor and autophagy to promote hepatocellular carcinoma metastasis by interacting with miR-455[J].DNA Cell Biol,2019,38(8):857-864.
[13] CHEN Q,XU H,ZHU J,et al.LncRNA MCM3AP-AS1 promotes breast cancer progression via modulating miR-28-5p/CENPF axis[J].Biomed Pharmacother,2020,128(1):110289-110299.
[14] KANG M,XIA P,HOU T,et al.MicroRNA-190b inhibits tumor cell proliferation and induces apoptosis by regulating Bcl-2 in U2OS osteosarcoma cells[J].Pharmazie,2017,72(5):279-282.
[15] LIU J,YAO L,ZHANG M,et al.Downregulation of LncRNA-XIST inhibited development of non-small cell lung cancer by activating miR-335/SOD2/ROS signal pathway mediated pyroptotic cell death[J].Aging(Albany NY),2019,11(18):7830-7846.
[16] WANG Q,WANG T,HU Y,et al.Circ-EIF4G3 promotes the development of gastric cancer by sponging miR-335[J].Pathol Res Pract,2019,215(9):152507-152517.
[17] XIE Y,DENG H,WEI R,et al.Overexpression of miR-335 inhibits the migration and invasion of osteosarcoma by targeting SNIP1[J].Int J Biol Macromol,2019,133(1):137-147.

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