Cancer stem cells (CSCs) are suggested to be responsible for high recurrence rate and metastasis of colorectal cancer (CRC). Identifying novel targets that can suppress CSCs proliferation and metastasis may provide novel approach to combat against CRC. In the present study, we examined the role of miR-148a in cisplatin-resistant CRC cells with enhanced stem cell marker expression and explored the underlying mechanisms. In this study, we used cisplatin to selectively enrich cisplatin-resistant CRC cells from SW480 cell line, and these selected cisplatin-resistant SW480 cells were with significantly enhanced expression of stem cell markers and increased chemoresistance. MicroRNA (miRNA) array and qRT-PCR assay identified the down-regulation of miR-148a in cisplatin-resistant SW480 cells. Overexpression of miR-148a suppressed expression of stem cell markers, inhibited sphere formation, invasion and migration, induced apoptosis, and reduced chemo-resistance in cisplatin-resistant SW480 cells. Bioinformatics prediction and luciferase reporter assay revealed that WNT10b was a downstream target of miR-148a, and overexpression of miR-148a suppressed WNT10b expression and β-catenin signaling activities. Enforced expression WNT10b attenuated the effects of miR-148a on cisplatin-resistant SW480 cells sphere formation, invasion and migration. Further study showed that overexpression of miR-148a also suppressed in vivo tumor growth, and WNT10b expression and β-catenin signaling activities in tumor tissues were suppressed by miR-148a overexpression. In the clinical samples, miR-184a was found to be down-regulated in CRC tissues, down-regulation of miR-148a predicted poor prognosis in CRC patients. In conclusion, our study for the first time enriched the cisplatin-resistant CRC cells with enhanced stem cell marker expression from sphere-forming and chemo-resistant SW480-derived tumor xenografts in immune-deficient mice, and miR-148a suppressed the expression of stem cell markers, increased chemo-sensitivity, cell invasion and migration at least partly via regulating WNT10b and β-catenin signaling pathway.

Hesperetin (HES) is a flavonoid that has been reported to exert protective effects against cardiac remodeling, lung fibrosis and hepatic fibrosis. However, reports on the effects and potential mechanisms of HES in renal fibrosis are limited. In the present study, a unilateral ureteric obstruction (UUO) mouse model and a transforming growth factor (TGF)‑β1‑activated normal rat kidney (NRK)‑52E cell model were established. HES was subsequently administered to these models to evaluate its anti‑fibrotic effects and potential underlying mechanisms of action. The results demonstrated that HES reduced obstruction‑induced renal injury and deposition of the extracellular matrix components collagen‑I and fibronectin in UUO mouse kidneys (P<0.05). Furthermore, HES treatment significantly suppressed EMT, as evidenced by decreased expression of α‑smooth muscle actin and E‑cadherin, (P<0.05). Additionally, HES inhibited the hedgehog signaling pathway in UUO mice and TGF‑β1‑treated NRK‑52E cells. The present findings indicate that HES treatment may inhibit EMT and renal fibrosis in vivo and in vitro by antagonizing the hedgehog signaling pathway.

This study investigated whether oxymatrine (OMT) treatment can ameliorate renal interstitial fibrosis in unilateral ureteral obstruction (UUO) mice model. Moreover, the potential mechanisms of such treatment were analyzed. Twenty-four C57/BL6 mice were randomly divided into three groups, namely sham group, vehicle plus unilateral ureteral obstruction (UUO)-treated group, and 100 mg/kg/d OMT plus UUO-treated group. All mice were euthanized seven days after surgery, and their kidneys were harvested. Renal injury, fibrosis, expression of proinflammatory cytokines, and the transforming growth factor-β1/Smads (TGF-β/Smads) and nuclear factor-kappa B (NF-κB)-signaling pathways were assessed. The results showed OMT significantly prevented kidney injury and fibrosis, as evidenced by decreased expression of collagen-1 and fibronectin. Furthermore, OMT administration inhibited the release of inflammatory factors including tumor necrosis factor-α, (TNF-α) interleukin-1β (IL-1β), and interleukin-6 (IL-6), as well as phosphorylated NF-κB p65. In addition, OMT blocked the activation of myofibroblasts by inhibiting the TGF-β/Smad3-signaling pathway. The findings indicate that OMT-attenuated renal fibrosis and inflammation, and this renoprotective effect may be ascribed to the inactivation of the TGF-β/Smad3 and NF-κB p65 pathways.

3,3′-Diindolylmethane (DIM), a natural acid condensation extracted from cruciferous plants, exhibits anti-fibrotic effects in hepatic and cardiac fibrosis models. The effects of DIM on renal fibrosis, however, are unclear. This study aimed to explore the protective effects of DIM on renal fibrosis. Unilateral ureteral obstruction (UUO) and transforming growth factor (TGF)-β1-stimulated normal rat kidney (NRK)-49F fibroblast cell mouse models were established. The models were then treated with DIM for the assessment of its anti-fibrotic effects and mechanisms. Results of HE and Masson staining showed that DIM reduced kidney injury and production of interstitial collagens fibrosis. CTS also inhibited expression of fibronectin, collagen-1 but retain E-cadherin in the UUO model. Furthermore, DIM suppressed local fibroblast activation, as evidenced by the suppressed expression of the myofibroblast markers α-SMA and vimentin in vivo and in vitro. In addition, DIM significantly inhibited the TGF-β1-induced proliferation of NRK49F cells in a time- and dose-dependent manner. DIM decreased Smad2/3 phosphorylation but increased Smad7 expression. Results suggested that DIM inhibits TGF-β/Smad2/3 signaling to attenuate renal interstitial fibrosis via inhibiting local fibroblast activation. This mechanism is likely related to Smad7 induction.

Colorectal cancer remains a significant challenge with high mortality rates. The aim of this study was to investigate the effect of targeting the microRNA-148a/WNT10b axis with the long non-coding RNA LINC00261 on the proliferation and apoptosis of colorectal cancer cells. In vitro, small interfering RNA-LINC00261 and microRNA-148 inhibitor sequences were synthesized and transfected into SW480 cells. The study groups included a control group, small interfering RNA negative control group, small interfering RNA group, small interfering RNA negative control + microRNA -inhibitor group, small interfering RNA + microRNA -inhibitor group, and small interfering RNA + microRNA-negative control group. The transfection efficiency and expression levels of LINC00261 and miR-148a were evaluated by quantitative reverse transcription polymerase chain reaction. Cell proliferation, apoptosis, cell cycle distribution, and protein expression levels of WNT10b and β-catenin were evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, flow cytometry, and Western blot, respectively. After small interfering RNA-LINC00261 transfection, a significant decrease in cell proliferation (p < 0.05) and an increase in apoptosis (p < 0.05) were observed, accompanied by cell cycle arrest in the G1 phase. Inhibition of LINC00261 by small interfering RNA resulted in increased microRNA-148a expression and decreased protein expression of WNT10b and β-catenin. However, the small interfering RNA + microRNA inhibitor group showed significantly increased levels of WNT10b and β-catenin protein expression. These results suggest that silencing of long non-coding RNA LINC00261 could potentially affect the proliferation of SW480 cells by regulating the micro RNA -148a/WNT10b axis.