The role of miR-144/GSPT1 axis in gastric cancer.

OBJECTIVE: To investigate the potential effects of miR-144/GSPT1 axis on the development of gastric cancer. PATIENTS AND METHODS: The expressions of GSPT1 (G1 to S Phase Transition 1) and miR-144 were detected in gastric cancer tissues and the adjacent normal tissues. We also explored the levels of GSPT1 and miR-144 in both normal gastric cell line (GES-1) and gastric cells (SGC7901). Luciferase assay was conducted to evaluate the interaction between miR-144 and GSPT1. The effects of the miR-144/GSPT1 axis on SGC7901 cells were determined via investigating cell proliferation, invasion and metastasis. RESULTS: miR-144 was found to be down-regulated in gastric cancer tissues while GSPT1 expression level was markedly increased. Bioinformatics analysis showed that GSPT1 was a direct target of miR-144. Luciferase assays confirmed our hypothesis. The subsequent experiments showed that miR-144 could promote cell proliferation, invasion and migration in gastric cancer cells via inhibiting GSPT1. CONCLUSIONS: We showed that miR-144/GSPT1 axis could be a potential therapeutic target in treatment of gastric cancer.

Expressions and correlation analysis of HIF-1α, survivin and VEGF in patients with hepatocarcinoma.

OBJECTIVE: To investigate the expressions of HIF-1α, surviving, and VEGF in patients with hepatocarcinoma as well as the correlation analysis among them. PATIENTS AND METHODS: 65 patients, who were admitted to our hospital and diagnosed as hepatocarcinoma from January 2014 to October 2015, were selected as hepatocarcinoma group, while 50 healthy cases that do not have hepatocarcinoma were selected as normal control group. The expression levels of HIF-1α, surviving, and VEGF in hepatocarcinoma tissues of hepatocarcinoma group and normal liver tissues of control group were detected by immunohistochemical (SP) staining method; then, the correlation among them was explored. The expression levels of HIF-1α, surviving, and VEGF protein in hepatocarcinoma tissues and corresponding normal tissues were detected by Western blot. RESULTS: The positive expression rate of HIF-1α, surviving, and VEGF in hepatocarcinoma tissues of hepatocarcinoma group was respectively 46.2%, 55.4%, and 61.5%, significantly higher than that in cancer adjacent normal liver tissues of control group which was 2%, 2%, and 2%, and the differences were statistically significant (p<0.05). The expressions of HIF-1α, surviving, and VEGF in hepatocarcinoma tissues of patients with hepatocarcinoma were correlated with clinical stage, tumor differentiation degree and extrahepatic metastasis (p<0.05), but were not related to gender and tumor size (p>0.05). By Spearman rank correlation analysis, it could be seen that HIF-1α expression was positively correlated with VEGF protein expression in hepatocarcinoma tissues (r=0.683, p<0.05). Survivin expression was positively correlated with VEGF protein expression (r=0.717, p<0.05). There was no significant correlation between HIF-1α expression and survivin expression (p>0.05). The relative quantitative value of HIF-1α, surviving, and VEGF in hepatocarcinoma tissues of hepatocarcinoma group was respectively 3.04±0.23, 2.26±0.31, and 2.57±0.36, significantly higher than that in cancer adjacent liver tissues of control group which was 1.07±0.17, 1.31±0.27, and 1.42±0.43, and the differences were statistically significant (p<0.05). From Western blot electrophoresis scanning, it could be seen that the expressions of HIF-1α, surviving, and VEGF in hepatocarcinoma tissues were higher than those in cancer adjacent normal liver tissues. CONCLUSIONS: The expressions of HIF-1α, surviving, and VEGF played important roles in the occurrence, invasion, and metastasis of hepatocarcinoma. In hepatocarcinoma tissues, HIF-1α, and survivin protein expression was positively correlated with VEGF expression, but survivin protein was not related to HIF-1α expression, which indicated that HIF-1α and survivin may inhibit the apoptosis of hepatocarcinoma cells and promote tumor angiogenesis by up-regulating the expression of VEGF protein, thus accelerating the occurrence and development of hepatocarcinoma.

[Screening for limonoid glucosides in Citrus grandis L. Osbeck by high performance liquid chromatography-electrospray ionization/mass spectrometry].

A screening method for limonoid glucosides in the peel of Citrus grandis L. Osbeck, utilizing high performance liquid chromatography (HPLC) with diode-array detection (DAD), interfaced to electrospray ionization-mass spectrometry (ESI-MS), has been developed. The methanol extract of citrus peel was preseparated with conventional polymer adsorption column and weak base anion ion-exchange column and then the eluate from the column was analysed by LC-ESI/MS. In this paper, the UV-Vis spectra and the mass spectra provide structural information without the necessity of isolating the individual compounds. Two major limonoid glucosides-obacunone glucoside (OG) and nomilin glucoside (NG) were identified in the methanol extract of the peel of Citrus grandis L. Osbeck. Then the two limonoid glucosides were taken as the target and isolated by means of preparative HPLC on a C18 reversed-phase column with an acidic acetonitrile-water mobile phase. The structures of OG and NG were further confirmed by NMR spectrometry. The results show that this method is rapid, convenient, sensitive and very useful for screening bioactive natural products.

[Research on the separation of limonoid glucosides by reversed-phase preparative high performance liquid chromatography].

Obacunone-17-beta-D-glucopyranoside (OG) was isolated from the seeds of Citrus Sinensis Osbeck by using solvent extraction, classical polymer adsorption column separation and weak base anion ion-exchange separation, OG was finally purified by C18 reversed-phase preparative high performance liquid chromatography and was identified by thin-layer chromatography. The purity of OG was analyzed by analytical reversed-phase HPLC. At last the structure of OG was determined by 1H and 13C nuclear magnetic resonance spectrometry (NMR). In this work, the conditions of the reversed-phase preparative HPLC technique to purify limonoid glucosides was optimized. The reversed-phase preparative HPLC on a C18 column with a mobile phase of acidic acetonitrile-water (about 0.2% TFA, V/V) at pH 3 enabled the baseline separation of limonoid glucosides in the extract. The results show that OG is the predominant limonoid glucoside in the seeds of Citrus Sinensis Osbeck and nomilin glucoside is the second one. The results also show that the classical polymer adsorption column separation and weak base anion ion-exchange separation are effective for purifying limonoid glucosides.