The Effect of miR-503-5p on the Proliferation, Invasion, Migration and Epithelial Interstitium of Cervical Cancer HeLa Cells via Targeting E2F3

RAN Wei, ZENG Yu-hua, MA Xiao-jie, LIAO Ping, LIU Xing-lan

Abstract

To investigate the effect of miR-503-5p on the proliferation, invasion, migration and epithelialization of cervical cancer HeLa cells via targeting E2F3.  Methods  Four ccervical cancer HeLa cells groups were set up including control group, mimic-NC group, miR-503-5p mimic group, E2F3 group, miR-503-5p mimic+E2F3 group (mimic+E2F3 group). The plasmids were separately or jointly transinfected into cervical cancer Hela cells of each group by Lipofectamine 2000, After transinfection, the target gene was predicted by gene prediction software, the targeting relationship was verified by fluorescein experiment, the expression of miR-503-5p and E2F3 was detected by RT-PCR, cell proliferation was detected by MTT assay, expression of Ki67, proliferating cell nuclear antigen (PCNA), E-cadherin and N-cadherin were detected by Western blot, cell invasion was detected by Transwell, and cell migration was detected by scratch test. Nude mice were divided into control group and miR-503-5p mimic group, and 0.2 mL of cervical cancer HeLa cell suspension transfected with mimic-NC or miR-503-5p mimic was injected subcutaneously into the ventral side of the right hind limb of nude mice. Thirty days post injection, the nude mice were sacrificed by cervical dislocation. The tumor weight was weighed by an electronic balance, and the expression of KI67 and Vimentin in the tumor tissue was detected by immunohistochemistry.  Results  The expression level of miR-503-5p in cervical cancer HeLa cells was down-regulated, miR-503-5p directly targeted E2F3 by binding with E2F3 at binding sites in the 3'UTR region. Over-expressing of miR-503-5p inhibited the expression of E2F3, significantly decreased cell growth rate and the expression level of Ki67 and PCNA, decreased the number of invasive cells, widened the scratches, reduced the healing rate, up-regulated the expression of E-cadherin and also down-regulated the expression of N-cadherin (P<0.01). Over-expressing of miR-503-5p significantly reduced the volume and weight of transplanted tumors, and decreased the proportion of positive Ki67 and Vimentin (P<0.01).  Conclusion  miR-503-5p inhibits the proliferation, invasion, migration and epithelialization of cervical cancer HeLa cells by targeting E2F3.

 

Keywords: miR-503-5p, E2F3, Cervical cancer HeLa cells

 

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References


DUENÃAS-GONZÃLEZ A， CAMPBELL S. Global strategies for the treatment of early-stage and advanced cervical cancer. Curr Opin Obstet Gynecol，2016，28(1): 11–17.

YAO X， TIAN Q， HE J， et al. miR-503 inhibits hepatocellular carcinoma cell growth via inhibition of insulin-like growth factor 1 receptor. Onco Targets Ther，2016，9(1): 3535–3544.

WU J， GAO F， XU T， et al. miR-503 suppresses the proliferation and metastasis of esophageal squamous cell carcinoma by triggering autophagy via PKA/MTOR signaling. Int J Oncol，2018，52(5): 1427–1442.

QIU T， ZHOU L， WANG T， et al. miR-503 regulates the resistance of non-small cell lung cancer cells to cisplatin by targeting BCL-2. Int J Mol Med，2013，32(3): 593–598.

GUPTA G， CHELLAPPAN D K， DE JAP T， et al. Tumor suppressor role of miR-503. Panminerva Med，2018，60(1): 17.

YIN Z L， WANG Y L， GE S F， et al. Reduced expression of miR-503 is associated with poor prognosis in cervical cancer. Eur Rev Med Pharmacol Sci，2015，19(21): 4081.

GONG C， LIU H， SONG R， et al. ATR-CHK1-E2F3 signaling transactivates human ribonucleotide reductase small subunit M2 for DNA repair induced by the chemical carcinogen MNNG. Biochim Biophys Acta，2016，1859(4): 612–626.

SU Z， YANG H， ZHAO M， et al. Microrna-92a promotes cell proliferation in cervical cancer via inhibiting p21 expression and promoting cell cycle progression. Oncol Res，2017，25(1): 137–145.

YEE G P C， SOUZA P D， KHACHIGIAN L M. Current and potential treatments for cervical cancer. Curr Cancer Drug Targets，2013，13(2): 205–220.

MELLONE M， RINALDI C， MASSIMI I， et al. Human papilloma virus-dependent HMGA1 expression is a relevant step in cervical carcinogenesis 1. Neoplasia，2008，10(8): 773–781.

PERALTA-ZARAGOZA O， BERMÚDEZ-MORALES V H， PÉREZ-PLASENCIA C， et al. Targeted treatments for cervical cancer: a review. Onco Targets Ther，2012，5: 315–328.

SUN Y， LI L， XING S， et al. miR-503-3p induces apoptosis of lung cancer cells by regulating p21 and CDK4 expression. Cancer Biomark， 2017，20(2): 1–12.

XU K， CHEN G， QIU Y， et al. miR-503-5p confers drug resistance by targeting PUMA in colorectal carcinoma. Oncotarget，2017，8(13): 21719–21732.

CHANG S W， YUE J， WANG B C， et al. miR-503 inhibits cell proliferation and induces apoptosis in colorectal cancer cells by targeting E2F3. Int J Clin Exp Pathol，2015，8(10): 12853.

GENG D， SONG X， NING F， et al. miR-34a inhibits viability and invasion of human papillomavirus-positive cervical cancer cells by targeting E2F3 and regulating survivin. Int J Gynecol Cancer，2015，25(4): 707–713.

GUO J， LIU X， WANG M. miR-503 suppresses tumor cell proliferation and metastasis by directly targeting RNF31 in prostate cancer. Biochem Biophys Res Commun，2015，464(4): 1302–1308.

LI B， LIU L， LI X， et al. miR-503 suppresses metastasis of hepatocellular carcinoma cell by targeting PRMT1. Biochem Biophys Res Commun， 2015，464(4): 982–987.

ZHAO Z， FAN X， JIANG L， et al. miR-503-3p promotes epithelialemesenchymal transition in breast cancer by directly targeting Smad2 and E-cadherin. J Genet Genomics，2017，44(2): 75–84.


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