Fibroblast growth factor-4 and hepatocyte growth factor induce differentiation of human umbilical cord blood-derived mesenchymal stem cells into hepatocytes

AIM： To investigate the differentiation of human umbilical cord blood （HUCB）-derived mesenchymal stem cells （MSCs） into hepatocytes by induction of fibroblast growth factor-4 （FGF-4） and hepatocyte growth factor （HGF）, and to find a new source of cell types for therapies of hepatic diseases. METHODS： MSCs were isolated by combining gradient density centrifugation with plastic adherence. When HUCB-derived MSCs reached 70% confluence, they were cultured in Iscove modified Dulbecco medium （IMDM） supplemented with 10 mL/L FBS, 20 ng/mL HGF and 10 ng/mL FGF-4. The medium was changed every 4 d and stored for albumin, alpha-fetoprotein （AFP） and urea assay. Expression of CK-18 was detected by immunocytochemistry. Glycogen storage in hepatocytes was determined by PAS staining. RESULTS： By combining gradient density centrifugation with plastic adherence, we could isolate MSCs from 25.6% of human umbilical cord blood. When MSCs were cultured with FGF-4 and HGF, approximately 63.6% of cells became small, round and epithelioid on d 28 by morphology. Compared with the control, the level of AFP increased significantly from d 12 to 18.20：1=1.16 μg/L （t = 2.884, P〈0.05） in MSCs cultured with FGF-4 and HGF, and was higher （54.28±3.11 μg/L） on d 28 （t = 13.493, P〈0.01）. Albumin increased significantly on d 16 （t = 6.68, P〈0.01） to 1.02±0.15 μg/mL, and to 3.63±0.30 μg/mL on d 28 （t = 11.748, P〈0.01）. Urea （4.72±1.03 μmol/L） was detected on d 20 （t = 4.272, P〈0.01）, and continued to increase to 10.28±1.06 μmol/L on d 28 （t = 9.276, P〈0.01）. Cells expressed CK-18 on d 16. Glycogen storage was observed on d 24. CONCLUSION： HUCB-derived MSCs can differentiate into hepatocytes by induction of F-GF-4 and HGF. HUCB derived MSCs are a new source of cell types for cell transplantation therapy of hepatic diseases.

Effect of abnormal MZF1 and FoxM1 expression on cell proliferation and apoptosis in osteosarcoma

Objective: To investigate the effect of abnormal MZF1 and FoxM1 expression on cell proliferation and apoptosis in osteosarcoma. Methods: A total of 88 patients with osteosarcoma who underwent surgical treatment in Zigong Third People's Hospital between March 2014 and June 2017 were selected, 88 cases each of intraoperative osteosarcoma tissue and adjacent normal tissue were collected as osteosarcoma group and adjacent normal tissue group respectively. Fluorescence quantitative PCR method was used to detect the expression of MZF1, FoxM1, proliferation genes and apoptosis genes, and Pearson test was used to evaluate the correlation of MZF1 and FoxM1 gene expression with the malignant biological behaviors of cancer cells in osteosarcoma. Results: MZF1 mRNA expression in osteosarcoma group was lower than that in adjacent tissue group;FoxM1 mRNA expression was higher than that in adjacent tissue group;proliferation genes Med19, IRX2 and RIPK4 mRNA expression were higher than those in adjacent tissue group whereas YM155 mRNA expression was lower than that in adjacent tissue group;apoptosis genes Survivin and bcl-2 mRNA expression were higher than those in adjacent tissue group whereas Caspase8 and bax mRNA expression were lower than those in adjacent tissue group. Pearson test showed that the abnormal expression of MZF1 and FoxM1 in osteosarcoma was directly correlated with the proliferation and apoptosis activity of cancer cells. Conclusion: MZF1 expression abnormally decreases whereas FoxM1 abnormally increases in osteosarcoma, and they are directly related to the proliferation and apoptosis activity of cancer cells.

Effect of Twist, Snail and YB-1 gene expression in cervical cancer tissue on cell invasion and epithelial-mesenchymal transition

Objective:To study the effect of Twist, Snail and YB-1 gene expression in cervical cancer tissue on cell invasion and epithelial-mesenchymal transition. Methods:Cervical cancer tissue samples and tissue samples adjacent to carcinoma were collected from 138 patients with radical operation for cervical cancer, fluorescence quantitative PCR method was used to detect the mRNA expression of Twist, Snail and YB-1 genes, cell invasion-related genes and epithelial-mesenchymal transition marker genes, the Pearson test was used to analyze the correlation of Twist, Snail and YB-1 gene mRNA expression in cervical cancer tissue with cell invasion and epithelial-mesenchymal transition.Results:Twist, Snail and YB-1 gene mRNA expression in cervical cancer tissue were higher than those in tissue adjacent to carcinoma, the invasion genes STAT3, YAP1, TUG1, FoxM1 and Rab11 mRNA expression were higher than those in tissue adjacent to carcinoma, and the epithelial-mesenchymal transition markers E-cadherin andβ-catenin gene mRNA expression were lower than those in tissue adjacent to carcinoma while vimentin gene mRNA expression was higher than that in tissue adjacent to carcinoma. Pearson test showed that Twist, Snail and YB-1 gene mRNA expression in cervical cancer tissue were directly correlated with cell invasion and epithelial-mesenchymal transition. Conclusion:Twist, Snail and YB-1 genes are highly expressed in cervical cancer tissue, and their abnormal expression directly leads to the increased tumor cell invasion activity and the aggravated epithelial-mesenchymal transition.