| Methods:(1) The expression of OPCML protein was detected by immunohistochemistry in gastric carcinoma tissues and adjacent tissues, and to evaluate the correlation of OPCML with clinicopathological characteristics and prognosis of gastric cancer.(2) The OPCML expression in human gastric cancer cell lines(7901, AGS, BGC823, MKN28, N87, KATO, SNU1) and normal gastric mucosa was assessed by RT-PCR.(3) The recombinant plasmid pc DNA3.1-OPCML was constructed and transfected into AGS cells, with pc DN A3.1-Vector as a control group, the expression levels of OPCML m RNA and protein were detected by RT-PCR and Western after transfection.(4) Using CCK8, colony formation assay, soft agar colony formation assay and scratch test to analysis the impact on the proliferation, colony formation and migration ability of the AGS after overexpression of OPCML.Results:(1) OPCML was low expression in gastric cancer, accounting for 68.6%; OPCML expression had a significant inverse correlation with depth of tumor invasion and tumor differentiation degreein in 118 cases of gastric carcinoma(p<0.05); OPCML expression, depth of tumor invasion, lymph node metastasis and distant metastasis were closely related to the prognosis of patients(p<0.05).(2) OPCML m RNA expression in gastric cancer cells was significantly lower than in normal gastric mucosa.(3) RT-PCR and Western showed that the OPCML expression was aberrantly increased in cells transfected with pc DNA3.1-OPCML.(4) CCK8 showed that after transfection of OPCML, the proliferation of AGS cells decreased significantly.(5) Colony formation assay showed that the overexpression of OPCML inhibition of the clone formation of AGS cell(quantity, volume). The colony numbers of transfection group and the control group are respectively(8.33±5.29)and(29.87±10.82), and the average difference of the cloning efficiency between transfection group and control group was 72%.(6) Soft agar colony formation assay showed that transfected and control group were forming colonies(23.33 ± 6.66) and(69.67±10.50), the relative difference between cloning efficiency was 67%, with a significant difference between the two(P <0.05).(7) Scratch test showed at 0 h, 24 h, 48 h, compared to the control group, the transfected group migration smaller and scratch width significantly larger.Conclusion:(1) OPCML was low expression or lack of expression in human gastric cancer tissue and gastric cancer cell lines; OPCML expression was significantly associated with infiltrating depth and differentiation degree in gastric cancer;OPCML expression was related to the prognosis of patients with gastric cancer.(2) OPCML has significant inhibitory effect on biological behavior of gastric cancer cell proliferation and migration. |
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