长链非编码RNA SNHG3对人乳腺癌细胞MCF-7增殖、迁移与侵袭的影响
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摘要
目的:探讨长链非编码RNA SNHG3对人乳腺癌细胞MCF-7增殖、迁移与侵袭的影响。方法:构建SNHG3过表达质粒,实验分别设置阴性对照组(pcDNA-3. 1+)与SNHG3基因过表达组(pcDNA-3. 1+/SNHG3)。将MCF-7细胞转染对照组质粒和SNHG3过表达质粒,采用实时定量PCR方法检测SNHG3 mRNA转录水平,Western blot检测MMP9及EMT相关蛋白质水平;集落形成实验检测MCF-7细胞增殖能力;划痕愈合实验检测MCF-7细胞横向迁移能力; Transwell小室实验检测MCF-7细胞纵向迁移能力及侵袭能力。结果:过表达SNHG3后,MCF-7细胞中SNHG3的mRNA水平显著增高(P <0. 001); MCF-7细胞的体外增殖能力明显增加(P <0. 01),迁移(P <0. 01)与侵袭能力(P <0. 001)也显著增强,实时定量PCR,Western blot结果显示SNHG3可激活EMT相关通路。结论:过表达SNHG3可能通过激活EMT通路促进乳腺癌MCF-7细胞的增殖,迁移与侵袭。
Objective: To investigate the effects of LncRNA SNHG3 on the proliferation,migration and invasion of human breast cancer MCF-7 cells. Methods: The recombiant plasmid of SNHG3 was constucted. The experiment was divided into two group: control group( transfected with pcDNA-3. 1 + plasmid) and treatment group( transfected with pcDNA-3. 1 +/SNHG3 plasmid). The mRNA level of SNHG3 was detected by qRTPCR; The mRNA and protein levels of MMP9,EMT-related makers were measured by qRT-PCR and Western blot; The colony formation assay was used to test the proliferation of MCF-7 cells; The wound-healing and Transwell chamber assay were taken to evaluate the migration and invasion of MCF-7 cells. Results: The expression of SNHG3 in MCF-7 cells was highly upregulated compared with control group( P < 0. 001);Furthermore,The mRNA and protein levels of MMP9 and EMT-related markers were increased; The migration and invasion abilities of SNHG3 overexpression group measured by wound healing assay and transwell chamber assay were augmented significantly. And the colony formation assay showed that proliferation of treatment group were increased remarkably. Conclusion: Overexpression of SNHG3 may promote the proliferation,migration and invasion of breast cancer MCF-7 cells by activating the EMT signaling pathway.
Objective: To investigate the effects of LncRNA SNHG3 on the proliferation,migration and invasion of human breast cancer MCF-7 cells. Methods: The recombiant plasmid of SNHG3 was constucted. The experiment was divided into two group: control group( transfected with pcDNA-3. 1 + plasmid) and treatment group( transfected with pcDNA-3. 1 +/SNHG3 plasmid). The mRNA level of SNHG3 was detected by qRTPCR; The mRNA and protein levels of MMP9,EMT-related makers were measured by qRT-PCR and Western blot; The colony formation assay was used to test the proliferation of MCF-7 cells; The wound-healing and Transwell chamber assay were taken to evaluate the migration and invasion of MCF-7 cells. Results: The expression of SNHG3 in MCF-7 cells was highly upregulated compared with control group( P < 0. 001);Furthermore,The mRNA and protein levels of MMP9 and EMT-related markers were increased; The migration and invasion abilities of SNHG3 overexpression group measured by wound healing assay and transwell chamber assay were augmented significantly. And the colony formation assay showed that proliferation of treatment group were increased remarkably. Conclusion: Overexpression of SNHG3 may promote the proliferation,migration and invasion of breast cancer MCF-7 cells by activating the EMT signaling pathway.
引文
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[14]Tang Y,Xiao G,Chen Y,et al. LncRNA MALAT1 promotes migration and invasion of non-small-cell lung cancer by targeting miR-206 and activating Akt/m TOR signaling. Anti-Cancer Drugs,2018,29(8):725-735.
[15]Wu W,Chen F,Cui X,et al. LncRNA NKILA suppresses TGF-β-induced epithelial-mesenchymal transition by blocking NF-κB signaling in breast cancer. International Journal of Cancer,2018,143(9):2213-2224.
[16]Cabibbo G,CraxìA. Epidemiology,risk factors and surveillance of hepatocellular carcinoma. European Review for Medical&Pharmacological Sciences,2010,14(4):352.
[17] Chen G G, Ho R L, Wong J, et al. Single nucleotide polymorphism in the promoter region of human alpha-fetoprotein(AFP)gene and its significance in hepatocellular carcinoma(HCC). Eur J Surg Oncol,2007,33(7):882-886.
[18] Li N,Zhan X,Zhan X. The lncRNA SNHG3 regulates energy metabolism of ovarian cancer by an analysis of mitochondrial proteomes. Gynecologic Oncology,2018,150(2):343-354.
[19]Hou Z,Xu X,Fu X,et al. HBx-related long non-coding RNA MALAT1 promotes cell metastasis via up-regulating LTBP3 in hepatocellular carcinoma. American Journal of Cancer Research,2017,7(4):845-856.
[20] Kotiyal S. Bhattacharya Breast cancer stem cells,EMT and therapeutic targets Biochem. Biophys. Res Commun,2014,453(1):112-116.
[21]May C D,Sphyris N,Evans K W,et al. Epithelial-mesenchymal transition and cancer stem cells:a dangerously dynamic duo in breast cancer progression. Breast Cancer Research,2011,13(1):202.
[2]Lin C,Yang L. Long noncoding RNA in cancer:wiring signaling circuitry. Trends in Cell Biology,2018,28(4):287-301.
[3]Terracciano D,Terreri S,De F N,et al. The role of a new class of long noncoding RNAs transcribed from ultraconserved regions in cancer. Biochim Biophys Acta,2017,1868(2):449.
[4] Ballantyne M D,Pinel K,Dakin R,et al. Smooth muscle enriched long noncoding RNA(SMILR)regulates cell proliferation. Circulation,2016,133(21):2050-2065.
[5] Sun T,Wong N. Transforming growth factor-β-induced long noncoding RNA promotes liver cancer metastasis via RNA-RNA crosstalk. Hepatology,2015,61(2):722-724.
[6] Ranzani V,Arrigoni A,Rossetti G,et al. Next-generation sequencing analysis of long noncoding RNAs in CD4+T cell differentiation. Methods Mol Biol,2017,1514:173-185.
[7]Padua D,Mahurkar-Joshi S,Law I K,et al. A long noncoding RNA signature for ulcerative colitis identifies IFNG-AS1 as an enhancer of inflammation. Am J Physiol Gastrointest Liver Physiol,2016,311(3):G446-G457.
[8]Jia P,Cai H,Liu X,et al. Long non-coding RNA H19 regulates glioma angiogenesis and the biological behavior of gliomaassociated endothelial cells by inhibiting microRNA-29a. Cancer Lett,2016,381(2):359-369.
[9]Yang F,Zhang H,Mei Y,et al. Reciprocal regulation of HIF-1αand lincRNA-p21 modulates the Warburg effect. Mol Cell,2014,53(1):88-100.
[10]Zhang T,Cao C,Wu D,et al. SNHG3 correlates with malignant status and poor prognosis in hepatocellular carcinoma. Tumor Biology,2016,37(2):2379-2385.
[11] Chan S,Friedrichs K,Noel D,et al. Prospective randomized trial of docetaxel versus doxorubicin in patients with metastatic breast cancer. Journal of Clinical Oncology,1999,17(8):2341-2354
[12]Kawaji H,Severin J,Lizio M,et al. Update of the FANTOM web resource:from mammalian transcriptional landscape to its dynamic regulation. Nucleic Acids Research,2009,10(4):R40.
[13]He H,Wei Z,Du F,et al. Transcription of HOTAIR is regulated by RhoC-MRTF-A-SRF signaling pathway in human breast cancer cells. Cellular Signalling,2017,31:87-95.
[14]Tang Y,Xiao G,Chen Y,et al. LncRNA MALAT1 promotes migration and invasion of non-small-cell lung cancer by targeting miR-206 and activating Akt/m TOR signaling. Anti-Cancer Drugs,2018,29(8):725-735.
[15]Wu W,Chen F,Cui X,et al. LncRNA NKILA suppresses TGF-β-induced epithelial-mesenchymal transition by blocking NF-κB signaling in breast cancer. International Journal of Cancer,2018,143(9):2213-2224.
[16]Cabibbo G,CraxìA. Epidemiology,risk factors and surveillance of hepatocellular carcinoma. European Review for Medical&Pharmacological Sciences,2010,14(4):352.
[17] Chen G G, Ho R L, Wong J, et al. Single nucleotide polymorphism in the promoter region of human alpha-fetoprotein(AFP)gene and its significance in hepatocellular carcinoma(HCC). Eur J Surg Oncol,2007,33(7):882-886.
[18] Li N,Zhan X,Zhan X. The lncRNA SNHG3 regulates energy metabolism of ovarian cancer by an analysis of mitochondrial proteomes. Gynecologic Oncology,2018,150(2):343-354.
[19]Hou Z,Xu X,Fu X,et al. HBx-related long non-coding RNA MALAT1 promotes cell metastasis via up-regulating LTBP3 in hepatocellular carcinoma. American Journal of Cancer Research,2017,7(4):845-856.
[20] Kotiyal S. Bhattacharya Breast cancer stem cells,EMT and therapeutic targets Biochem. Biophys. Res Commun,2014,453(1):112-116.
[21]May C D,Sphyris N,Evans K W,et al. Epithelial-mesenchymal transition and cancer stem cells:a dangerously dynamic duo in breast cancer progression. Breast Cancer Research,2011,13(1):202.
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