The Long Non-Coding RNA NEAT1 Promotes Gastric Cancer Cell Proliferation and Invasion by Regulating miR-103a/ STAMBPL1 Axis.

BACKGROUND: Gastric cancer (GC) is a common malignancy with high morbidity. Long non-coding RNAs (LncRNAs) have been demonstrated to be critical post-transcriptional regulators in tumorigenesis. This study aimed to investigate the effect of LncRNA NEAT1 on the proliferation and metastasis of GC. MATERIAL AND METHODS: The expression of LncRNA NEAT1 was examined in clinical samples and GC cell lines. GC cell lines (SGC-7901 and BGC-823) and human normal gastric epithelial cell line (GES-1) were employed. The correlation between NEAT1, miR-103a and STAMBPL1 was determined by luciferase reporter assay. Cell viability was determined by CCK8 assay. Cell invasion capacity was examined by Transwell assay. The protein level of STAMBPL1 was analyzed by western blotting. RESULTS: LncRNA NEAT1 was found to be up-regulated in GC cell lines. Further studies identified LncRNA NEAT1 as a direct target of miR-103a. Moreover, NEAT1 knockdown and miR-103a overexpression inhibited cell proliferation and cell invasion. NEAT1 knockdown and miR-103a overexpression also decreased STAMBPL1 levels. CONCLUSION: Our study indicated that LncRNA NEAT1 was up-regulated in GC cells and tissues. NEAT1 was targeted and inhibited by miR-103a and acted as an oncogene, which promoted the malignant behavior of GC cells. This regulatory effect of NEAT1 may be associated with STAMBPL1. Therefore, NEAT1 could be used as a biomarker for predicting the progression of GC.

STAMBPL1 knockdown has antitumour effects on gastric cancer biological activities.

The present study aimed to investigate the effects and mechanisms of STAM binding protein-like 1 (STAMBPL1) knockdown in the suppression of gastric cancer activities. Pathological data and STAMBPL1 protein expression were analysed in 36 patients with gastric cancer, including 24 stage I-II and 12 stage III-IV patients, by haematoxylin and eosin staining and immunohistochemistry. In vitro cell experiments were performed to measure AGS cell proliferation, apoptosis, invasion and migration by MTT, Celigo cell count, flow cytometry, Transwell and wound healing assays following STAMBPL1 knockdown. The relative protein expression levels were evaluated by western blotting. When compared with the adjacent normal tissues, STAMBPL1 protein expression in the gastric cancer tissues with increasing stages was significantly upregulated (P<0.01 or P<0.001). STAMBPL1 gene expression was not identified to be significantly different between AGS and MGC80-3 gastric cancer cells (P>0.05). Following STAMBPL1 knockdown by short hairpin RNA (sh)STAMBPL1, cell proliferation was significantly suppressed, the cell apoptosis rate was significantly upregulated, and the numbers of invasive AGS cells and the AGS wound healing rate were significantly decreased (P<0.01 and P<0.001, respectively), compared with those in the shControl group. Additionally, STAMBPL1 and NF-κB protein expression levels were significantly downregulated in the shSTAMBPL1 group (P<0.001, respectively). STAMBPL1 may be oncogenic in gastric cancer, and STAMBPL1 knockdown may suppress gastric cancer development.

[Expression of FOXC-2 and YB-1 in Gastric Carcinoma and Its Role in Invasion and Metastasis].

OBJECTIVE: To investigate the expression of FOXC-2,YB-1 and related proteins and their influences on development,invasion and metastases in gastric carcinoma. METHODS: A total of 193 tissue samples were collected,including 50 cases of normal gastric mucosa, 50 cases of gastric mucosal intraepithelial neoplasia and 93 cases of primary gastric carcinoma. The 93 cases of primary gastric carcinoma included 74 cases of positive lymph node metastasis tissues, 19 cases of nonmetastasis tissues and 33 cases of distant metastasis tissues. Immuohistochemistry was used to detect the expression and distribution of FOXC-2,YB-1,E-cadherin, Vimentin and MMP-2 in normal and intraepithelial neoplasia,gastric carcinoma,positive lymph node metastasis tissues and distant metastasis tissues. RESULTS: The expressions of FOXC-2,YB-1,Vimentin and MMP-2 in gastric carcinoma were significantly higher than those in normal and intraepithelial neoplasia while the expression of E-cadherin was significantly lower (P<0.05).The expressions of FOXC-2,YB-1 were significantly correlated with low expression of E-cadherin and high expression of Vimentin and MMP-2 (P<0.05). The expression of FOXC-2 protein was significantly correlated with TNM stage,lymph node metastasis and distant metastases (P<0.05).The expression of YB-1 protein was significantly correlated with TNM stage,differentiation degree,invasion depth,lymph node metastasis and distant metastasis (P<0.05). The expression of MMP-2 protein was closely related to the degree of differentiation,invasion depth,lymph node metastasis and distant metastasis (P<0.05). CONCLUSION: FOXC-2, YB-1 may be related to the occurrence, development, invasion and metastasis of gastric carcinoma. The possible mechanism is to promote the invasion and metastasis of cancer cells by activating the epithelial mesenchymal transition process and up regulating the expression of MMP-2.

Ginsenoside-Rh2 Inhibits Proliferation and Induces Apoptosis of Human Gastric Cancer SGC-7901 Side Population Cells.

OBJECTIVES: To observed the effects of ginsenoside -Rh2 (GS-Rh2) on proliferation and apoptosis of side population (SP) human gastric cancer SGC-7901 cells. MATERIALS AND METHODS: SGC-7901 SP and Non-SP cells were sorted by flow cytometry and assessed using the cck-8 method. Expression of apoptosis-related proteins Bax and Bcl-2 of SP before and after the intervention was determined by Western-blotting. RESULTS: It was found that the proliferation of SP was significantly faster than that of NSP (<0.05). In addition, GS-Rh2 inhibited proliferation of gastric cancer SP cells, induced cell cycle arrest and cell apoptosis, and changed the expression of BAX/Bcl-2 proteins in a time-dependent and concentration-dependent manner (<0.05). CONCLUSIONS: With increase of GS-Rh2 dose, GS-Rh2 gradually inhibit the proliferation of SGC-7901 SP cells, which have high proliferation rate, through G1/G0 phase arrest, followed by apoptosis which involves the up-regulation of Bax and the down-regulation of Bcl-2.

[Effect of Chinese herbal therapy on T-lymphocytes of IgA nephropathy patients: a clinical observation].

OBJECTIVE: To observe the effect of Chinese herbal therapy on T-lymphocyte subsets in patients with IgA nephropathy (IgAN). METHODS: Totally 36 inpatients and outpatients at Department of Nephropathy, Xiyuan Hospital, China Academy of Chinese Medical Sciences, from June 2011 to June 2013 were recruited in the treatment group, while 20 volunteers were recruited as the healthy control group. Patients in the IgAN group only took Chinese herbal decoctions by syndrome typing for 3 months (except those accompanied with hypertension additionally took antihypertensive agents such as angiotensin-converting enzyme inhibitor and/or dihydropyridines calcium antagonist). No intervention was performed in the healthy control group. The values of Th1, Th2, and CD4+ CD25+ Treg, and red blood cell number in urine were detected using flow cytometry before and after treatment. 24 h urine protein was detected using inmmunoturbidimetry. RESULTS: Compared with the healthy control group, the CD4+ CD25+ Treg level obviously decreased in the IgAN group, showing statistical difference (P < 0.01). In the IgAN group, Th1, 24 h urine protein, and urine red blood cell counts were obviously lower after treatment, showing statistical difference when compared with before treatment (all P < 0.05). CONCLUSION: Chinese herbal therapy could reduce urine erythrocyte number and 24 h urine protein of IgAN patients, and down-regulating Th1 expression might be its mechanism.

[Effect of Sijunzi decoction on the proliferation of side population cells of human gastric cancer cell line].

OBJECTIVE: To observe the proliferation changes of the side population of gastric cancer cell line SGC-7901 cells (SP), the non-side population (NSP) cells, and unsorted cells (Total) after intervened by Sijunzi Decoction (SD) containing serum. METHODS: Sixteen pure bred New Zealand rabbits were equally divided into the normal control group, the low dose SD group (at the daily dose of 7 mL/kg), the middle dose SD group (at the daily dose of 14 mL/kg), and the high dose SD group (at the daily dose of 28 mL/kg) according to the random digit table. Rabbits' serum was extracted after equal volume of corresponding medication was given by gastrogavage twice daily for 2 consecutive weeks. The drug serum was identified using high performance liquid chromatography. SP cells of SGC-7901 were detected using flow cytometry, SP and NSP cells were screened. The proliferation curve of SP, NSP, and Total cells were detected with CCK-8 assay. Changes of their proliferation were also observed. RESULTS: Ginsenoside Rg1, an effective ingredient in SD was detected in prepared drug serum. The proliferation of SGC-7901 SP cells was significantly higher than that of NSP cells and Total cells (P < 0.05). Drug serum on gastric cancer cell line SGC-7901 SP, NSP, and Total cells could inhibit their proliferation, but its inhibition on SP cells' proliferation was significantly lower than on NSP and Total cells (P < 0.05). CONCLUSIONS: SD could significantly inhibit the proliferation of gastric cancer cell line SGC-7901 SP, NSP, and Total cells. But there exist obvious difference in the inhibition among the three groups.

[Regulatory effects of yiqi zishen granule on helper T-lymphocyte subsets in IgA nephropathy patients].

OBJECTIVE: To study the pathogenesis of IgA nephropathy (IgAN) and the effect of Yiqi Zishen Granule (YZG) on the helper T-lymphocyte subsets Th1/Th2 in IgAN patients. METHODS: Intracellular cytokines secreted by Th1/Th2 were measured using flow cytometry in 24 healthy subjects and 31 IgAN patients treated by YZG before and after treatment. RESULTS: Level of interferon-gamma (IFN-gamma) was lower (P < 0.05) and levels of interleukin 4 (IL-4) and interleukin 10 (IL-10) were higher (P < 0.01) in the IgAN patients than those in the healthy controls, showing imbalance of Th1/Th2 in the IgAN patients. After YZG treatment, the level of IFN-gamma increased and IL-10 level decreased (P < 0.01). CONCLUSION: Imbalance of Th1/Th2 exists in IgAN patients. YZG has a regulatory effect on imbalance of Th1/Th2 in IgAN patients.