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Objective:To examine the effect of FAM83D knockdown on proliferation, survival ability and invasion of human esophageal squamous cell carcinoma after X-ray radiation, and explore the mechanism.Methods:The expression of FAM83D, E-cadherin and vimentin in tumor tissues was detected in 69 cases of esophageal squamous cell cancer by using immunohistochemical method. The siRNA based on the sequences of the FAM83D mRNA were synthesized to transfect into the cultured ECA109 cells as FAM83D shRNA group. The effect of silencing FAM83D gene was evaluated to determine the protein levels of FAM83D in the human oesophageal squamous cell carcinoma ECA109 and KYSE30 cells using western blotting. MTS, clone formation, and Transwell assay were employed to examine the proliferation, survival ability and invasion of ECA109 and KYSE30 cells in vitro, respectively. We used flow cytometry assay to analyze distribution of cell apoptosis in different groups. Western blotting was used to examine the expression of cell metastasis-related molecules and apoptosis-related protein. Results:The strong expression rates of FAM83D, E-cadherin, and vimentin were 55%(38/69), 36%(25/69) and 61%(42/69) in the tumor tissues, respectively. FAM83D protein expression was significantly and negatively correlated with the expression of E-cadherin ( r=-0.350, P<0.01), and positively with the expression of vimentin ( r=0.470, P<0.01). Western blotting results demonstrated that silencing FAM83D gene significantly reduced the FAM83D protein expression ( P<0.01). MTS data demonstrated that FAM83D knockdown after irradiation significantly inhibited the proliferation of esophageal squamous cell carcinoma ECA109 and KYSE30 cells ( P<0.05). The data from the clone formation assay revealed that the radiosensitivity was increased after downragulation of FAM83D expression ( P<0.01). In addition, the invasive abilities of oesophageal carcinoma cells transfected with FAM83D shRNA after irradiation were significantly inhibited compared with those of the NC group ( P<0.01), followed by the downregulation of N-cadherin, vimentin, Snail, p-Akt and p-GSK-3β expression, and the upregulation of E-cadherin expression ( P<0.01). The apoptosis rate of tumor cells in FAM83D shRNA group after irradiation was markedly increased ( P<0.01), followed by a decrease of Bcl-2 and Mcl-1 expression and an increase of Cleaved caspase-3 expression ( P<0.01). Conclusions:FAM83D expressions was found to be closely related to the invasion and development of ESCC. Furthermore, siRNA interference technology inhibited the expression of FAM83D gene in oesophageal squamous cell carcinoma cells, reduced the proliferation, invasion of cells, induced cell apoptosis, and increased radiosensitivity, which may be associated with regulating the epithelial-mesenchymaltransition via Snail/Akt/GSK-3β signaling pathways.
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Bladder cancer is the most common malignant tumor in urogenital system. Bladder carcinoma has the biological characteristics of easy recurrence and metastasis. The cancer stem cells theory explained the mechanisms of bladder cancer recurrence and metastasis. Epithelial-mesenchymal transition ( EMT) could promote tumor cells to acquire stem cell characteristics, while the tumor cells with stem cell characteristics have correspondingly high expression of EMT biomarkers. Long noncoding RNA ( lncRNA) promotes tumor cells to acquire and maintain stem cell characteristics through regulating EMT process. CSCs, EMT and lncRNA are closely related to the genesis and development of tumors, and participate in the regulation process of tumor proliferation, invasion, me-tastasis and so on. It will provide new insights in targeting therapy of tumor metastasis and recurrence through illuminating the correla-tion between lncRNA on EMT of bladder cancer stem cells ( BCSCs) This article makes a review on the latest research progress about the mechanism of lncRNA regulating EMT of BCSCs.
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Objective To observe the effects of different concentrations of SB203580, the inhibitor of P38MAPK, in process of high glucose (GS)-induced renal tubular epithelial-myofibroblast transdifferentiation (TEMT). Methods The cultured human renal tubular epithelial cells (HK-2) were divided into control group (5.5 mmol/L GS), GS (30 mmol/L GS) group and different concentrations of SB203580 (30 mmol/L GS +5, 10, 20 and 30 μmol/L SB203580) groups. The treat?ments were for 48 hours. MTT assay was used to observe cell proliferation. The median inhibiting concentration (IC50) was cal?culated. Western blot assay was used to detect the expressions of P38MAPK, P-P38MAPK andα-smooth muscle actin (α-SMA) in control group, high-glucose group and S30 group. The expression ofα-SMA was also detected by the method of im?munofluorescence. Results 1.Compared with control group, there was no significant inhitory effect on proliferation rate in DMSO group (P>0.05). There were increased HK-2 cells in high glucose group and S5group (P0.05). 3. Compared with control group, the expression ofα-SMA was signifi?cantly increased in high glucose group and S30 group (P<0.05). Compared with high glucose group, the expression of α-SMA was significantly decreased in S30 group (P < 0.05). Conclusion The 30 mmol/L GS can lead to TEMT in HK-2 cells. The more suitable inhibitory concentration of SB203580 in the process of TEMT is 30μmol/L. SB203580 can slow down the process of TEMT by inhibiting P38MAPK activation and inhibiting proliferation and the expression ofα-SAM s of HK-2 cells.
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Objective To explore the mechanism of the inflammatory factors to increase the reanl inferstitial fibrosis by stndying on epithelial-to-mesenchymal transition induced by IL-1?.Methods Mice primary tubular epithelial cells were Cultured in vitro,then stimulated by IL-1? for 3,4,5 days respectively,from which the protein was extracted and the protein expression of ?-SMA and Vimentin was detected with western bloHing;while the cells were stimnlated by IL-1? for 12hs,36hs and 60hs respectively,from which then total RNA was extracted and the mRNA expression of ?-SMA and Vimentin was detected by ranscription-polymerase chain reaction analyses(RT-PCR).those without IL-1?stimulating were regarded as control group.Results The protein levels of ?-SMA,Vimentin in tubular epithelial cells stimulated with IL-1?for 3 days or 4 days were significantly higher than that in normal group cells.Those stimulated for 5 days,the protein levels of ?-SMA,Vimentin in tubular epithelial cells increased very high.The mRNA expression of ?-SMA and Vimentin increased significantly in 12hs in tubular epithelial cells after IL-1?stimulating as compared with normal group cells and presented time-depend.Conclusions IL-1? plays the role of promoting renal interstitial fibrosis by inducing the transdifferentiation of renal tubular epithelial cells and reducing the proliferation cells probably.