Colorectal cancer cells induce the formation of cancer-associated fibroblasts by activating the ERK signaling pathway in fibroblasts / 南方医科大学学报

OBJECTIVE@#To investigate the mechanism by which conditioned medium of colorectal cancer cells promotes the formation of cancer-associated fibroblasts (CAFs).@*METHODS@#Normal human colorectal fibroblasts (CCD-18Co cells) in logarithmic growth phase were treated with the conditioned media of colorectal cancer HCT116 cells (HCT116-CM) or Caco-2 cells (Caco-2-CM) alone or in combination with 300 nmol/L ERK inhibitor SCH772984. The expression levels of CAFs-related molecular markers were detected in the treated cells with real-time quantitative PCR (RT- qPCR) and immunofluorescence assay, and the changes in cell proliferation, colony formation and migration were assessed with RTCA, colony formation and wound healing assays; Western blotting was performed to detect the activated signaling pathways in the fibroblasts and the changes in CAFs formation after blocking of the signaling pathway.@*RESULTS@#HCT116-CM and Caco-2-CM significantly upregulated mRNA expression levels of CAFs markers (including α-SMA, FAP, FN and TGF-β) in CCD-18Co cells, and strongly promoted fibroblast transformation into CAFs (P < 0.05). The two conditioned media also promoted the proliferation, colony formation and migration of CCD-18Co cells (P < 0.05) and significantly increased the levels of α-SMA protein and ERK phosphorylation in the cells (P < 0.05). The ERK inhibitor SCH772984 obviously inhibited the expression of α-SMA and the transformation of CCD-18Co cells into CAFs induced by the conditioned medium of colorectal cancer cells (P < 0.05).@*CONCLUSION@#Colorectal cancer cells may induce the formation of colorectal CAFs by activating the ERK pathway in the fibroblasts.

A pilot study of molecular mechanism of salmon milt DNA (SMD) retards mouse insenescence / 中国组织工程研究

Aim To iuvestigate the effect and the mechanism of salmonmilt DNA (SMD) on age-related involutions in mouse thymus. MethodsFemale BALB/c of 10 months were divided randomly into three groupsaccording to their weights: high dosage group 333.33 mg/(kg @ d), lowdosage group 166. 67 mg/(kg @ d) and control group 0 mg/(kg @ d) .After five weeks, with Image-Pro Plus (version. 4.0) software, the thymusindexes and the thymoctytes in the thymus section were measured, as wellas the thymus cortex thickness. All the data were analyzed by SAS statisticsoftware. Mieroarray technique was applied to screen the gene fragments,which were differently expressed between the high dosage group and thecontrol group, together with RT-PCR to further confirm some of them.Results No significant differences of the variables including bodyweight, thymus weight and thymus indexes among the three groups werefound (F ＜ 3.0 and P ＞ 0.05, respectively). The thymocytes quanti-ties of thymus cortex and medulla in the high dosage group were significantlyhigher than those of the control group [cortex D(H, C) = 9.46, P ＜ 0.01;medulla t( H.C) = 2.53, P ＜ 0.05]. The thymus cortex thicknesses of bothSMD supplement groups were significantly higher than that of the control group[cortex D(L,C)=3.65, P＞ 0.05; medulla t(L, C)=0.8, P＞ 0.05] .112differently expressed gene fragments were isolated. Furthermore, we foundthe fragments with the logged number of U23789, X80232 and Aw209102were highly expressed in the high dosage group when RT-PCR techniquewas used. Conclusion SMD may reverse the age-related involutions inmouse thymus via up-regulation the expression of proliferation related genesand development and differentiation related genes simultaneously.