Lnc ⁃ NEAT1 regulates the proliferation,migration and invasion of melanoma B16 cells through the miR⁃29c⁃3p/CSPG4 signaling axis
CSTR:
Author:
Affiliation:

Clc Number:

R329.2

Fund Project:

  • Article
  • |
  • Figures
  • |
  • Metrics
  • |
  • Reference
  • |
  • Related
  • |
  • Cited by
  • |
  • Materials
  • |
  • Comments
    Abstract:

    Objective:To investigate the regulatory mechanism of long non-coding RNA-nuclear enriched transcript 1(lnc-NEAT1) on the proliferation,migration and invasion of melanoma(MM)cells. Methods:MM cell lines(A375,A875,M14,B16)and human epidermal melanocytes(HEMa - LP)were cultured in vitro,real -time quantitative PCR(RT - qPCR)was performed to determine the mRNA expression of lnc-NEAT1,microRNA-29c-3p(miR-29c-3p)and chondroitin sulfate proteoglycan 4(CSPG4)in cells. B16 cells at logarithmic growth phase were taken and separated into control group,si-NC group,si-NEAT1 group,si-NEAT1+inhibitor-NC group, and si-NEAT1+miR-29c-3p inhibitor group. Lipofectamine 3000 was applied to transfect the corresponding plasmids into cells. RT-qPCR was performed to determine the transfection efficiency. MTT method was used to determine cell proliferation. Transwell assay was performed to determine cell migration and invasion abilities. Western blotting was performed to determine the expression of CSPG4 and proteins related to the proliferation,migration and invasion. Dual-luciferase reporter gene assay was performed to determine miR-29c-3p targeting relationship with lnc-NEAT1 and CSPG4. Nude mice xenograft experiment was performed to explore the effect of lnc-NEAT1 knockdown on the growth of MM cells in vivo. Results:The mRNA levels of lnc-NEAT1 and CSPG4 in MM cell lines were obviously higher than those in HEMa-LP cells,and the levels of miR-29c-3p were obviously lower than those in HEMa-LP cells(all P < 0.05). Knockdown of lnc -NEAT1 obviously increased miR - 29c -3p expression,decreased CSPG4 mRNA and protein levels,inhibited cell proliferation,migration and invasion,and decreased Ki-67,N-cadherin and Vimentin protein levels as well as increased E-cadherin protein level(all P < 0.05). Down - regulation of miR -29c -3p expression obviously increased CSPG4 mRNA and protein levels,and attenuated the inhibitory effects of lnc-NEAT1 knockdown on MM cell proliferation,migration and invasion(P < 0.05). The results of dual-luciferase reporter gene assay showed that after the transfection of miR-29c-3p mimic,the luciferase activities of NEAT1-WT and CSPG4-WT in cells were obviously decreased(P < 0.05). Nude mouse xenograft experiments showed that knockdown of lnc -NEAT1 obviously inhibited the growth of xenografts in vivo,while inhibition of miR-29c-3p was able to obviously attenuate the inhibitory effect of lnc-NEAT1 knockdown on the growth of xenografts(all P < 0.05). Conclusion:Lnc-NEAT1 may promote the proliferation,migration and invasion of MM cells by regulating the miR-29c-3p/CSPG4 axis

    Reference
    Related
    Cited by
Get Citation

谢奇,樊鑫梅,韩永红,陶娟,刘旭,刘家秀. lnc⁃NEAT1通过miR⁃29c⁃3p/CSPG4信号轴调节黑色素瘤B16细胞的增殖、迁移和侵袭[J].南京医科大学学报(自然科学版英文版),2023,43(4):492-501.

Copy
Share
Article Metrics
  • Abstract:
  • PDF:
  • HTML:
  • Cited by:
History
  • Received:
  • Revised:
  • Adopted:
  • Online: April 23,2023
  • Published:
Article QR Code