ABSTRACT
Stanniocalcin-1 (STC1) is secreted by the variety of tissues having a major role in the regulation of calcium ions in the involuting mammary gland. The present work aims to sequence and structural characterization as well as expression profiling of STC1 gene in buffalo. Polymorphism identified in the 3-untranslated region (UTR) was analysed by polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) genotyping in riverine and swamp buffaloes. Expression profiling of STC1 was performed in different lactation stages of mammary gland and peripheral blood mononuclear cells to study the impact of 3'-UTR polymorphism on its expression. Different polymorphic sites were detected in the entire coding and noncoding regions of riverine and swamp buffaloes, including two INDELs. An identified polymorphic nucleotide locus A324G, having target sites for two miRNAs, namely bta-miR-2382 and bta-miR-1343, reported in cattle, was genotyped by PCR-RFLP to reveal variable allelic distribution among swamp and riverine buffaloes. Gene expression profiling across buffalo mammary tissues representing different lactation stages showed maximum expression of the STC1 gene in the involuting mammary gland. Ruminants’ specific genetic variation has been observed in STC1 and its implication in buffalo mammary gland involution as well as coregulation of gene expression throughmiRNA binding in the 3'-UTR is suggested.
ABSTRACT
Objective To investigate effect of stanniocalcin-1 (STC1) gene on the proliferation,apoptosis and radiotherapy sensitivity of non-small cell lung cancer.Methods The STC1 siRNA (STC 1-siRNA) and the non-interfering siRNA (negative control group) were transfected into the human lung cancer A549 cells by LipofectamineTM2000,and the blank control group was established.The expression level of STC1 protein was detected after transfection for 48 h by Western blotting.Clone forming test was adopted to detect the proliferation of A549 cells after STC1-siRNA and irradiation treatment.CCK8 assay was performed to detect the cell viability after treatment with STC1-siRNA and STC1-siRNA+8 Gy.The cell apoptosis was detected by flow cytometry.The expression levels of Ki67,Bax,STAT3 and p-STAT3 proteins were quantitatively measured by Western blotting.Results The expression level of STC1 protein in the A549 cells transfected with STC1-siRNA was significantly down-regulated than that in the blank control group (P＜ 0.05).Compared with the blank control group,the sensitization ratio was significantly enhanced after STC1-siRNA transfection.Compared with the blank control group,the cell viability and the expression levels of Ki67 and p-STAT3 protein were significantly decreased,whereas the apoptosis rate and the expression of Bax protein were significantly increased in the STC1-siRNA group.Compared with the STC1-siRNA group,the cell viability and the expression levels of Ki67 and p-STAT3 proteins were significantly decreased,whereas the cell apoptosis rate and the expression of Bax protein were remarkably increased in the STC1-siRNA+ 8 Gy group (all P＜0.05).Conclusion Inhibition of STC1 gene expression can enhance the radiotherapy sensitivity and down-regulate the STAT3 signaling pathway in non-small cell lung cancer.
ABSTRACT
Objective: To observe the effect of STC-1 gene expression was inhibited on the apoptosis, IL-1β and TNF-α expression and JAK2/STAT3 signal in esophageal cancer cells. Methods: Compared with normal human esophageal squamous epithelial cells Het-1 A, STC-1 expression was detected in human esophageal squamous cell carcinoma KYSE170, Eca109, TE1 and TE10 cells by RTPCR and Western blot; the siRNA sequence that the synthesized STC-1 and the si NRA sequence without interference were transfected into Eca109 cells, which were labeled as STC-1-siRNA group and NC group, and the blank control group was set, cells were transfected for 48 h, the expression of STC-1 were detected by RT-PCR and Western blot. Eca109 cell viability and apoptosis rate were detected by CCK8 and flow cytometry. IL-1β and TNF-α expression were detected by RT-PCR; the expression of Ki67, p53, p-JAK2 and p-STAT3 protein were detected by Western blot. Results: Compared with Het-1 A cells, expression of STC-1 mRNA and protein in KYSE170, Eca109, TE1 and TE10 cells were increased significantly ( P＜0. 05); compared with the control group, STC-1 expression was decreased significantly in STC-1-siRNA group, cell viability was decreased significantly in STC-1-siRNA group, the apoptosis rate was increased significantly in STC-1-siRNA group; IL-1β, TNF-α, Ki67, p-JAK2 and p-STAT3 expression were decreased significantly in STC-1-siRNA group, p53 expression was increased significantly in STC-1-siRNA group ( P＜0. 05). Conclusion: STC-1 was highly expressed in esophageal cancer cells; inhibiting of STC-1 expression could significantly reduce the activity of cancer cells, and increase apoptosis rate, this effect may be related to the inhibition of JAK2/STAT3 signaling pathways and inflammatory factors as IL-1β and TNF-α.