Expression Of Neuropilin-1 In Gastric Cancer Cell AGS And Its Function

Objectives To transfect the neuropilin 1(NRP-1) small interference RNA(si RNA) into human gastric cancer cell line AGS by Lip2000,and silence the neuropilin 1 m RNA, Then observe the NRP- 1 effects on gastric cancer cell lines series of biological behavior, For the study of an NRP- 1 as the targets for new targeted drug treatment for cancer of the gastric to provide necessary theoretical basisMethods Regular training gastric cancer cell lines AGS, in strict accordance with the instructions of si RNA.First, to transfect the neuropilin 1(NRP-1) small interference RNA(si RNA) into human gastric cancer cell line AGS by Lip2000 AGS cells, After 48 hours, observe the transfection efficiency of AGS cell lines in fluorescence microscop, transfection efficiency of more than 60% can treat successful transfection. By Lip-2000 to transfecti different substances into gastric cancer cell lines AGS, Gastric cancer cell lines can be divided into three different groups, namely the transfection group(transfection si RNA), meaningless sequences(transfection of si RNA nonsense sequence) and blank control group(transfection buffer PBS). First of all, to draw cell growth curve, by real-time fluorescent quantitative PCR detection of NRP-1 in three groups, respectively relative m RNA expression; Immunocytochemistry method in three groups of cells was detected in the expression of ki-67 protein; Determined by MTT respectively determine the proliferation of three groups; Flow cytometry instrument experiments the cell cycle of the three groups respectively; Transwell Chambers experiments invasion and migration ability of the three groups.Results 1 Rt-PCR results show that: after transfect NRP1-siRNA into AGS cells, the transfection group NRP- 1 m RNA expression level was significantly lower, statistically significant difference(P < 0.05).2 To identify according to the result of immunocytochemistry staining: the transfection group compared with control group, the relative expression of ki-67 protein significantly lower, statistically significant difference(P < 0.05).3 Determined by MTT method and cell growth curve showed that: the proliferation ability of transfection group was significantly reduced, statistically significant difference(P < 0.05).4 Flow cytometry test results showed that the transfection group growth inhibition, stagnation in the Gl period, a significant increase in the control group s, the difference statistically significant(P < 0.05).5 Migrating chamber experiments show that the transfection group compared with control groups, migratory ability decreased significantly, the difference statistically significant(P < 0.05).6 Chamber invasive experiments show that the transfection group compared with control groups, a significant reduction in the invasive ability, statistically significant difference(P< 0.05).Conclusions 1 Application of RNAi technology, design and synthesis of siRNA for NRP-1, can reduce the expression of NRP-1 m RNA effectively in AGS cells after transfection.2 After transfected NRP1-si RNA into AGS gastric cancer cells, the expression of ki-67 protein was significantly decreased, and cell proliferation capacity decreased obviously, the stagnation of cells in G1 phase.3 After transfected NRP1-si RNA into AGS gastric cancer cells,a significant reduction in the invasive and migrant ability of malignant biological behavior is restrained.4 NRP-1 in gastric cancer cells plays an important role in gastric cancer cells, can cause a series of biological behaviour, may be a candidate genes of gastric cancer gene therapy, provide theoretical foundation for gastric cancer therapeutic targets, has potential clinical application prospects.