Mir-124Inhibits Gastric Carcinoma Growth And Invasion By Targeting SPHK1

Objective: To investigate the expression of the miR-124in gastric carcinomaand their clinical significances.Methods: We selected176cases of gastric cancer tissue samples,adjacent tissues as anormal control.Hybridization in situ was used to detect the miR-124expression inTissue Array of the gastric carcinoma tissues and adjacent comparison tissues. Cellgrowth and cell invasion were detected by MTT, cell wound healing assay,transwellinvasion assay. miR-124targeting SPHK1was detected by Western blot andluciferase reporter vector.Results: Hybridization in situ showed that the positive expression rate of themiR-124was18.18%in176cases of gastric carcinoma tissues, and86.36%in adjacentcomparison tissues, the groups were significant different(P<0.01)； Thedown-regulation of miR-124expression was associated with advanced clinical stageand the depth of infiltration, the groups were significant different (P<0.01)；theexpression of miR-124was associated with the differentiation of gastric cancer, thepoorer the differentiation, the louwer the expression, the groups were significantdifferent(P<0.01)；the expression of miR-124was lower with lymph node metastasisthan without lymph node metastasis in gastric carcinoma, the groups were significantdifferent(P<0.05); Transfection of miR-124mimics for12h,24h,48h respectively,MTT assay showed that the OD value was (0.152±0.003),(0.216±0.001),(0.242±0.020), compared with the control group (0.316±0.010),(0.429±0.002)(0.517±0.007), the groups were significant different (P <0.05), and it is a time-dependentmanner; Transfection of miR-124mimics for12h,24h respectively, cell woundhealing assay showed that the wound healing rate was (0.243±0.005)(0.341±0.026),compared with the control group (0.399±0.015)(0.745±0.013), it can significantlyslow down the scratch wound healing of the MGC-803cell, the groups weresignificant different (P <0.05); Transfection of miR-124mimics for24h, transwell invasion assay showed that the number of cells through the basement membrane was(43±5), compared with the control group (108±17), it can significantly slow downthe invasion of MGC-803cell, the groups were significant different (P <0.05);Luciferase reporter vector system confirmed that SPHK1(sphingosine kinase,Sphingosine kinase1, SPHK1) was a target gene of miR-124.; Western blot showedthat the expressions of SPHK1,MMP-2and MMP-9protein were inhibited bymiR-124.Conclusions:1. miR-124expression level was downregulated in gastric carcinoma and had a closenegative correlation to advanced clinical stage and with lymph node metastasis.2. SPHK1is the target gene of miR-124.3. miR-124can inhibit gastric carcinoma cell growth and ivasion by inhibiting theexpressions of SPHK1, MMP-2and MMP-9.