Rab25 gene down-regulates snail expression and inhibits epithelial mesenchymal transformation of human breast cancer MDA-MB-231 cells / 中华内分泌外科杂志

Objective:To probe into Rab25 Gene’s Effect on TGF-β inhibition of proliferation, invasion and epithelial mesenchymal transformation (EMT) of breast cancer MDA-MB-231 cells and explore its molecular mechanism.Methods:The experiment was divided into three groups: control group,TGF-β Group and si-Rab25 group. TGF-β induced MDA-MB-231 cell model of EMT was built. CCK-8 assay was used to detect cell proliferation. Transwell assay was used to detect the ability of cell invasion and migration.Western blot was used to detect the changes of related proteins in each group.Results:After stimulating MDA-MB-231 cells with TGF-β, Rab25 gene was highly expressed. Compared with TGF-β group (57.48±%3.62%), the migration ability and invasion ability of cells in si-Rab25 group (33.49%±2.93%) decreased by 41.7%, with a significant difference ( P<0.05). Compared with TGF-β group (153.21%±4.17%), the proliferation ability of cells in si-Rab25 group (115.32%±5.69%) decreased by 24.73%, with a significant difference ( P<0.05). The expression of MDA-MB-231 fine EMT related protein in si-Rab25 group was significantly different from that in TGF-β group ( P<0.05). The expression of p-AKT and Snail protein in si-Rab25 group was significantly lower than that in TGF-β group ( P<0.05) . Conclusions:Rab25 gene is highly expressed in MDA-MB-231 cells. Silencing Rab25 gene can activate AKT signal pathway, inhibit Snail protein expression, regulate EMT related protein expression, and inhibit EMT transformation.

Research progress of Rab25 in tumor metastasis / 医学研究生学报

Metastasis is the main reason that affects the survival and death of cancer patients. In recent years, it has been found that Rab25 is closely related to tumor invasion and metastasis. Rab25 has dual role in tumor metastasis, which can be used as oncogene or tumor suppressor gene. In most tumor types, Rab25 is an oncogene, and only in a few tumors, Rab25 shows tumor suppressor function. However, the dual action mechanism and signal pathway of Rab25 are not very clear. This article summarizes the related research on the role of Rab25 in tumor metastasis.

Rab25 Deficiency Perturbs Epidermal Differentiation and Skin Barrier Function in Mice

Rab25, a member of the Rab11 small GTPase family, is central to achieving cellular polarity in epithelial tissues. Rab25 is highly expressed in epithelial cells of various tissues including breast, vagina, cervix, the gastrointestinal tract, and skin. Rab25 plays key roles in tumorigenesis, mainly by regulating epithelial differentiation and proliferation. However, its role in skin physiology is relatively unknown. In this study, we demonstrated that Rab25 knock-out (KO) mice show a skin barrier dysfunction with high trans-epidermal water loss and low cutaneous hydration. To examine this observation, we investigated the histology and epidermal differentiation markers of the skin in Rab25 KO mice. Rab25 KO increased cell proliferation at the basal layer of epidermis, whereas the supra-basal layer remained unaffected. Ceramide, which is a critical lipid component for skin barrier function, was not altered by Rab25 KO in its distribution or amount, as determined by immunohistochemistry. Notably, levels of epidermal differentiation markers, including loricrin, involucrin, and keratins (5, 14, 1, and 10) increased prominently in Rab25 KO mice. In line with this, depletion of Rab25 with single hairpin RNA increased the expression of differentiation markers in a human keratinocyte cell line, HaCaT. Transcriptomic analysis of the skin revealed increased expression of genes associated with skin development, epidermal development, and keratinocyte differentiation in Rab25 KO mice. Collectively, these results suggested that Rab25 is involved in the regulation of epidermal differentiation and proliferation.

The effects of Leptin in proliferation of MCF-7 breast cancer cells / 实用医学杂志

Objective To investigate the effects of leptin on proliferation of the breast carcinoma MCF-7 cells in terms of protein and nRNA of Rab-25 and C-myc.Methods MCF-7 cells were treated with leptin and normal saline (NS) as control.The 490 nm OD values of cells were read by MTT and the protein and mRNA expressions of Rab-25 and C-myc were measured by western blot and RT-PCR methods in two groups.Results Compared with control group,leptin can promote the proliferation of MCF-7 cells in a dose and time dependent manner within certain limit and up-regulate the protein expressions of Rab-25 and C-myc in the breast carcinoma MCF-7 cells (P ＜ 0.05).However,leptin also can up-regulate the mRNA expressions of Rab-25 but without significance (P =0.05).On the contrary,leptin can also up-regulate the mRNA expressions of C-myc (P ＜ 0.05).Conclusion Leptin promotes breast cancer cells proliferation,which may be related to up-regulate the protein and mRNA expressions of Rab-25 and C-myc.It could provide theoretical foundation and new target of gene therapy in the clinical treatment of breast cancer.

Study on apoptosis of human ovarian cancer cell induced by siRNA on Rab25 / 肿瘤研究与临床

Objective To study the effect of siRNA of Rab25 on apoptosis induction in ovarian carci- noma cell.Methods According to Rab25 mRNA sequence in the Genebank,the plasmids expressing siRNA against Rab25 were constructed and stably transected into A2780 cells,MTY method were applied to measure cell growth and proliferation.Apoptosis rate and cell cycle phase distribution of A2780 cells were measured by flow cytometry(FCM).Apoptosis was confirmed by agarose gel electrophoresis(AGE)of DNA.Results Cells transected with the plasmids expressing siRNA targeting Rab25 gene effectively decreased cell growth, proliferation;blocked A2780 cells in the G_1 phase of cell cycle and induced cell apoptosis.A typical DNA ladder pattern appeared on AGE.Conclusion Rab25 gene siRNA can inhibit growth and induce apoptosis in ovarian carcinoma cell line,which will facilitate further studies of Rab25 function and its application in the treatment of ovarian cancer.