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Effect of Grp78 on Invasion and Metastasis of Human Gastric Carcinoma SGC7901 Cells in Vitro and Theirs Mechanism

Author: YanXiaoLi
Tutor: GaoQing
School: Chongqing Medical University
Course: Internal Medicine
Keywords: Glucose-regulated protein 78 Gastric cancer BAPTA-AM Shift Epithelial mesenchymal transition
CLC: R735.2
Type: Master's thesis
Year: 2011
Downloads: 47
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Abstract


Objective: To investigate the calcium chelator BAPTA-AM and calcium ionophore A23187 gastric cancer cell line SGC7901 GRP78 expression level, analysis of the the GRP78 expression changes of gastric cancer cell invasion and metastasis, and to explore its possible mechanism. METHODS: Cultured human gastric cancer SGC7901 cell line and divided into BAPTA-AM treatment group and the control group and A23187 treatment groups, using RT-PCR and Western-blot to detect the expression of GRP78 expression at the mRNA and protein levels in vitro invasion assay and wound healing assay changes in cell invasion and metastasis ability, using immunocytochemistry, Western-blot detection of cells in each group E-cadherin and N-cadherin expression, analysis of GRP78 in gastric cancer cell metastasis in the mechanism of action. Results: A23187 group, the control group, 20μmol/LBAPTA-AM, 40μmol/LBAPTA-AM group, and of GRP78-mRNA 60μmol/LBAPTA-AM groups with the internal control of β-actin gray value ratio: 1.070 ± 0.005, respectively , 1.011 ± 0.012,0.701 ± 0.009,0.543 ± 0.009 and 0.348 ± 0.013 of GRP78-mRNA expression levels of the A23187 group compared with the control group increased (P lt; 0.01), compared with the control expression levels in three groups of BAPTA-AM group decreased (P lt; 0.01); A23187 group, the control group and 40μmol/LBAPTA-AM group GRP78 protein with the gray values ??of the internal control β-actin ratios were: 0.911 ± 0.004,0.823 ± 0.007,0.647 ± 0.015, GRP78 The expression levels of the protein in the A23187 group compared with the control group increased (P lt; 0.01), in the expression of BAPTA-AM group were decreased (P lt; 0.01). 2. A23187 48 hours scratches degree of healing higher than the control group (79.68% ± 0.91% vs 57.32% ± 0.80%, P lt; 0.01), the mean number of cells per field of view than the control group increased (89 ± 2 vs 59 ± 2, P lt; 0.01); BAPTA-AM group 48 hours scratches degree of healing lowering compared with the control group (39.92% ± 0.79% vs 57.32% ± 0.80%, P lt; 0.01), the mean number of cells per field of view than the decrease in the control group (43 ± 1 vs 59 ± 2, P lt; 0.01). A23187 invasion and metastasis compared with the control group enhanced invasion and metastasis of BAPTA-AM group compared with the control group decreased. 3 immunocytochemistry, Western blot Experimental results show that the level of calcium ionosphere A23187 group of E-cadherin expression compared with control group decreased (0.106 ± 0.003 vs 0.122 ± 0.003, P lt; 0.01; 0.495 ± 0.010 vs 0.709 ± 0.008, P lt ; 0.01), the level of expression of the E-cadherin BAPTA-AM group compared with the control group increased (0.133 ± 0.003 vs 0.122 ± 0.003, P lt; 0.01; 0.915 ± 0.017 vs 0.709 ± 0.008, P lt; 0.01); A23187 Group N-cadherin expression levels compared with the control group increased (0.143 ± 0.003 vs 0.131 ± 0.003, P lt; 0.01; 0.541 ± 0.009 vs 0.344 ± 0.006, P lt; 0.01), calcium BAPTA-AM group N-stick protein expression levels compared with the control group decreased (0.104 ± 0.004 vs 0.131 ± 0.003, P lt; 0.01; 0.210 ± 0.004 vs 0.344 ± 0.006, P lt; 0.01). Conclusion: BAPTA-AM down GRP78 expression in gastric cancer cells SGC7901 A23187 can induce its expression. GRP78 expression level can affect the invasion and metastasis of gastric cancer cells, and its mechanism may be related to the GRP78 change the expression of E-cadherin and N-cadherin thus affecting the level of the tumor epithelial mesenchymal transformation.

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CLC: > Medicine, health > Oncology > Gastrointestinal Cancer > Gastric neoplasms
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