Abstract
BACKGROUND: APPL1 is highly expressed in gastric cancer tissues and it is positively correlated with TNM stage, depth of invasion, and lymph node metastasis of gastric cancer, but the specific mechanism of APPL1 on gastric cancer cell proliferation has not been defined. In our study, we prepared the gastric carcinoma cell line SGC-7901/APPL1 via plasmid transfection technique, then investigated the change of gastric carcinoma cells' proliferation ability, aimed to investigate the regulatory mechanism of APPL1 on gastric cancer cell proliferation.
METHODS The gastric carcinoma cell strain SGC-7901 was transfected by APPL1 gene plasmid as the experimental group; by blank plasmid as the control group. The expression of APPL1 on the experimental group was detected by Western-blot technique. The change of the experimental group cells ' proliferation ability was detected by MTT assay, The change of the experimental group cell cycle was detected by PI staining, The change of the experimental group cell cyclin proteins were detected by Western-blot technique.
RESULTS By the Western- blot technique, the expression level of APPL1 protein in the experimental group cell strain was up-regulated effectively(P<0.01). Meanwhile, the proliferation ability of the experimental group increased significantly was detected by MTT assay(P<0.05), By the PI staining, the percentage of the experimental group cell cycle in G1 phase was decreased(P<0.01), in S phase was increased(P<0.01). By the Western-bolt technique, the expression of Cyclin D1(P<0.05) was up-regulated(P<0.05) and p16, p27 was down-regulated(P<0.05), Cyclin E、p21、p57 were not changed(P>0.05).
CONCLUSION Through the plasmid transfection technique, the gastric carcinoma cell line that could overexpress APPL1 protein were prepared successfully and whose proliferation ability were enhanced.