Expression of miR-101-3p in Gastric Cancer and Its Effects on Invasion, Metastasis, and Angiogenesis of Gastric Cancer Cells by Targeting STC-1 Gene to Regulate PI3K/AKT Signaling Pathway / 肿瘤防治研究

Objective To explore the expression of miR-101-3p in gastric cancer and its mechanism on the invasion, metastasis, and angiogenesis of gastric cancer cells by targeting the STC-1 gene to regulate the PI3K/AKT signaling pathway. Methods qRT-PCR was used to detect the expression of miR-101-3p and STC-1 mRNA in gastric cancer tissues and BGC-823 cell and analyze the relationship between miR-101-3p expression and patients' clinical pathological factors. The cells were transfected with miRNA mimics and plasmids separately or in combination with LipofectamineTM 2000. TargetScanHuman prediction and dual-luciferase assay were used to verify the targeted regulation of miR-101-3p on STC-1. The effect and possible mechanism of miR-101-3p targeting the STC-1 gene on the invasion, metastasis, and angiogenesis of cancer cells were verified by scratch test, Transwell chamber test, Matrigel in vitro tube forming test, and Western blot assay. The development of the transplanted tumor was detected by nude mouse tumorigenicity test. Results The expression of STC-1 in gastric cancer tissues was higher than that in normal tissues. Compared with normal gastric tissues and GES-1 cells, miR-101-3p was down-regulated, and STC-1 mRNA was up-regulated in gastric cancer tissues and BGC-823 cell. The level of miR-101-3p was negatively correlated with the level of STC-1, and significantly correlated with the degree of tumor differentiation, TNM stage, and lymph node metastasis (P < 0.05). miR-101-3p directly targeted STC-1. The overexpression of miR-101-3p inhibited STC-1 expression and downregulated the expression of p-PI3K/PI3K, p-AKT/AKT, MMP-2, MMP-9, VEGF, and Ang2, consequently, inhibited tumor cell invasion, metastasis, and angiogenesis and reduced the size and weight of the transplanted tumors (P < 0.05). Conclusion miR-101-3p is down-regulated in gastric cancer and can target the STC-1 gene to regulate the PI3K/AKT signaling pathway and inhibit the invasion, metastasis, and angiogenesis of BGC-823 gastric cancer cells and the development of transplanted tumors in vivo.

Expression and activity identification of recombinant human stanniocalcin 1 / 军事医学

Objective To obtain recombinant human stanniocalcin 1 ( STC1 ) with biological activity in Escheri.coli cells expression.Methods The gene was cloned into pET32b( +) vector by fused with thioredoxin and His tag .E.coli BL21(DE3) competent cells were transfomed by the recombinant vector .After renaturation, the fusion protein was digested with thrombin and intact STC1 protein was purified from the digested protein using Ni ion affinity chromatography .Recombi-nant humanSTC1 protein was confirmed by Western blot analysis using goat anti-STC1 antibody.The biological activity of STC1 in rat was assayed using standard method for assessment of renal function .Results The recombinant human STC 1 fu-sion protein is successfully expressed in Escherichia coli, the fusion protein was purified by affinity chromatography from the inclusion body and renaturated .Intact hSTC1 protein was released by thrombin digestion and purified by Ni ion affinity col-umn.The intact STC1 proteins was confirmed by Western blot analysis .Rat bioassay revealed that STC1 boosted phosphate reabsorption.Conclusion Recombinant STC1 protein was successfully expressed and has native biological activities .This protein could be used as an antigen for the preparation of monoclonal antibody against humanSTC 1.

Stanniocalcin-1 protects bovine intestinal epithelial cells from oxidative stress-induced damage

Chronic enteritis can produce an excess of reactive oxygen species resulting in cellular damage. Stanniocalcin-1(STC-1) reportedly possesses anti-oxidative activity, the aim of this study was to define more clearly the direct contribution of STC-1 to anti-oxidative stress in cattle. In this study, primary intestinal epithelial cells (IECs) were exposed to hydrogen peroxide (H2O2) for different time intervals to mimic chronic enteritis-induced cellular damage. Prior to treatment with 200 microM H2O2, the cells were transfected with a recombinant plasmid for 48 h to over-express STC-1. Acridine orange/ethidium bromide (AO/EB) double staining and trypan blue exclusion assays were then performed to measure cell viability and apoptosis of the cells, respectively. The expression of STC-1 and apoptosis-related proteins in the cells was monitored by real-time PCR and Western blotting. The results indicated that both STC-1 mRNA and protein expression levels positively correlated with the duration of H2O2 treatment. H2O2 damaged the bovine IECs in a time-dependent manner, and this effect was attenuated by STC-1 over-expression. Furthermore, over-expression of STC-1 up-regulated Bcl-2 protein expression and slightly down-regulated caspase-3 production in the damaged cells. Findings from this study suggested that STC-1 plays a protective role in intestinal cells through an antioxidant mechanism.

Role of stanniocalcin 1 in brain injury of coal-burning-borne fluorosis rats / 中国地方病学杂志

Objective To observe the change of stanniocalcin 1 (STC1) and calcium content in brain of coal-burning-borne fluorosis rats,and to explore the role of STC1 in brain injury of coal-burning-borne fluorosis.Methods Twenty four male SD rats were randomly divided into control,low,medium,and high fluoride groups according to body mass.Control group was fed conventional rat chow(fluorinated 1.3 mg/kg),and low,medium and high fluoride groups fed with fluorinated feed(20.0,40.0,60.0 mg/kg).All rats were given distilled water and feed ad libitum.One hundred and eighty days after modeling,STC1 protein and gene expression in the brain tissue of rats were detected using immunohistochemistry and RT-PCR and calcium content of brain tissue was detected.Results The cell positive rates of STC1 in low,medium,high fluoride groups [(48.10 + 2.11)％,(54.90 ± 1.73)％,(79.30 ± 3.71)％] were significantly higher than that of the control group[(24.70 + 3.53)％,all P ＜ 0.05],the cell positive rate of high fluoride group was significantly higher than that of the low and medium fluoride groups (all P ＜ 0.05).The STC1 mRNA expression of low,medium and high fluoride groups (0.58 ± 0.09,0.85 ± 0.17,1.75 ± 0.04) were significantly higher than that in the control group(0.37 ± 0.12,all P＜ 0.05),the STC1 mRNA expressions of high fluoride group was significantly higher than that of the low and medium fluoride groups (all P ＜ 0.05).The brain cortex calcium ion concentrations of low,medium and high fluoride groups[(138.62 + 4.19),(167.43 + 6.57),(189.45 + 3.72)nmol/L] were significantly higher than that in the control group [(101.47 + 9.46)nmol/L,all P ＜ 0.05],the brain cortex calcium ion concentrations of high fluoride group was significantly higher than that of the low and medium fluoride groups(all P ＜ 0.05),and the medium fluoride groups was higher than the low groups (P ＜ 0.05).Conclusion STC 1 may be involved in brain damage of coal-burning-borne fluorosis rats through regulating calcium balance.

Expression of hSTC-1 gene in peripheral blood and its correlation with micrometastases of breast cancer / 肿瘤

Objective: To detect the expression of human stanniocalcin-1 (hSTC-1) mRNA in peripheral blood and tumor tissues of breast cancer patients, determine micrometastases in peripheral blood, and study the relationship between the micrometastases and malignant degree of breast cancer. Methods: The expression of hSTC-1 mRNA in peripheral blood and breast cancer tissues was detected by RT-PCR. The expression of estrogen receptor (ER), progesterone receptor (PR), and vascular endothelial growth factor (VEGF) in breast cancer tissues were detected by immunohistochemical method. Results: Expression of hSTC-1 mRNA was identified in breast cancer tissues from 28 of 30 patients (93.3%) and in peripheral blood from 19 of 51 patients (37.3%). Expression of hSTC-1 mRNA was not detected in the peripheral blood from 17 women without breast cancer. Expression of hSTC-1 mRNA in the peripheral blood significantly correlated with multiple clinicopathological factors (P < 0.05), including tumor size, metastasis of lymph nodes, TNM stage, and the expression of ER and VEGF. Conclusion: The expression of hSTC-1 mRNA in the peripheral blood of breast cancer patients suggests the occurrence of micrometastasis. It significantly correlates with multiple clinicopathological factors which determined the prognosis of breast cancer. So hSTC-1 is proposed as a new marker for detecting micrometastasis of breast cancer in the peripheral blood.

Detection and its significance of human Stanniocalcin-1 mRNA in the peripheral blood from patients with colorectal cancer / 中华消化杂志

Objective To study the relationship between the expression of human Stanniocalcin-1(hSTC-1) mRNA in the peripheral blood from patients with colorectal cancer and its malignant behavior. Methods RT-PCR was used to detect hSTC-1 mRNA in the peripheral blood from 57 patients with colorectal cancer. The peripheral blood from 14 patients with gastrointestinal inflammatory diseases, 15 healthy volunteers and 5 pregnant women were served as controls. Results The positive rate of hSTC-1 mRNA in 57 patients with colorectal cancer was 49.12% (28/57), and the mRNA expression of hSTC-1 was significantly related with the clinical stage of colon cancer (P