• Asian-Australasian Journal of Animal Sciences
• /
• 제33권10호
• /
• pp.1674-1682
• /
• 2020

Objective: This study aimed to elucidate the effect of miR-140 on the proliferation of porcine fetal fibroblasts (PFFs) and identify the target genes of miR-140 in PFFs. Methods: In this study, bioinformatics software was used to predict and verify target genes of miR-140. Quantitative polymerase chain reaction and western blot were used to detect the relationship between miR-140 and its target genes in PFFs. Dual luciferase reporter gene assays were performed to assess the interactions among miR-140, type 1 insulin-like growth factor receptor (IGF1R), and SRY-box 4 (SOX4). The effect of miR-140 on the proliferation of PFFs was measured by CCK-8 when PFFs were transfected with a miR-140 mimic or inhibitor. The transcription factor SOX4 binding to promoter of IGF1R was detected by chromatin immunoprecipitation assay (ChIP). Results: miR-140 directly targeted IGF1R and inhibited proliferation of PFFs. Meanwhile, miR-140 targeted transcription factor SOX4 that binds to promoter of porcine IGF1R to indirectly inhibit the expression of IGF1R. In addition, miR-140 inhibitor promoted PFFs proliferation, which is abrogated by SOX4 or IGF1R knockdown. Conclusion: miR-140 inhibited PFFs proliferation by directly targeting IGF1R and indirectly inhibiting IGF1R expression via SOX4, which play an important role in the development of porcine fetal.

• Molecules and Cells
• /
• 제24권3호
• /
• pp.323-328
• /
• 2007

SOX (Sry-related HMG box) family proteins, which have an evolutionarily conserved DNA binding domain, have crucial roles in cell differentiation. However, their target genes remain enigmatic. Some members of the SOX family may have roles in regulation of cell proliferation. We established stable NT2/D1 cell lines overexpressing SOX15 (SOX15-NT2/D1), and a modified 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay showed that the SOX15-NT2/D1 cells exhibited significantly slower growth than the controls. Flow cytometry analysis revealed that an increased fraction of the SOX15-NT2/D1 cells were in G1-G0. In addition, a microarray analysis identified 26 genes that were up-regulated in the SOX15-NT2/D1 cells, but none that were down-regulated genes. Among the up-regulated genes, IGFBP5, S100A4, ID2, FABP5, MTSS1, PDCD4 have been shown to be related to cell proliferation and/or the cell cycle.

• 생명과학회지
• /
• 제29권8호
• /
• pp.895-903
• /
• 2019

이 연구는 단일 세포로 분리된 유도만능줄기세포(induced pluripotent stem cells, iPSCs)에 anoikis 세포사멸을 억제할 수 있는 Rho-associated protein kinase (ROCK)의 억제제를 처리하여 iPSCs의 세포 생존성을 향상하고자 하였다. Episomal plasmid 방법으로 확립된 iPSCs를 단일세포로 분리한 후, ROCK 억제제 Y-27632 dihydrochloride (Y-27632)를 0 uM, 0.5 uM, 1 uM, 2.5 uM, 5 uM, 7.5 uM 및 10 uM 농도별로 5주일 동안 각각 처리하였을 때, 5 uM 이상의 농도에서 세포의 생존율이 유의적으로 향상되었고, 10 uM의 Y-27632을 0일, 1일, 2일, 3일, 4일 및 5일 동안 처리하였을 때, Y-27632의 노출 기간이 길어질수록 세포의 생존율이 유의적으로 향상되는 것을 관찰하였다. 그러나, Y-27632의 노출 후, iPSCs의 형태학적 분화가 관찰되어 10 uM의 Y-27632에서 5일 동안 iPSCs에 처리 한 후, 줄기세포학적인 특성을 비교 조사하였다. 우선, octamer-binding transcription factor 4 (OCT-4), homeobox protein NANOG (NONOG) 및 SRY-box 2 (SOX-2) 줄기세포 특이 유전자의 발현은 Y-27632를 처리한 실험군은 Y-27632를 처리하지 않은 대조군에서 서로 유의적인 차이를 나타내지 않았다. 또한, Y-27632를 처리한 실험군은 Y-27632를 처리하지 않은 대조군과 비교하여 telomerase 활성과 이것의 활성과 관련된 telomerase reverse transcriptase (TERT) 및 telomerase RNA component (TERC)의 유전자 발현에는 유의적인 차이가 없었다. 이상의 결과로 보아, iPSCs에 Y-27632를 처리하였을 때, iPSCs의 줄기세포의 특정을 유지하면서 anoikis에 의한 세포사멸을 감소시켜 세포 생존율이 증가한다는 것을 알 수 있었다.