Inhibition Of Nanog Expression By RNA Interference In Human Gastic Cancer Cell Line SGC-7901

Objective: To construct and select out the recombinant plasmidsshRNA-Nanog which could inhibite the expression of Nanog gene by RNAinterference technology, and transfect them into human gastric cancer cellline SGC-7901, then detect the expression of Nanog in SGC-7901andexamine the proliferation, cell cycle, apoptosis, migration and invasion ofSGC-7901, and it lays a foundation for genetic diagnosis and treatment ofgastric cancer.Methods:(1)Construct the interference plasmids pshRNA-Nanog andtransfect them into SGC-7901by proliferation. To detect the expression ofNanog gene by fluorescence microscopy, detect the inhibition efficiency byRT-PCR and Western blot, and select out the most efficient recombinantplasmid.(2)Transfer the most efficient recombinant plasmid shRNA-Nanoginto SGC-7901by proliferation and examine the effect on proliferation byCCK-8test, cell cycle and apoptosis by flow cytometry, migration andinvasion capacity of SGC-7901by transwell tests.Results:(1)The recombinant plasmid shRNA-Nanog were constructed and transfect into SGC-7901by proliferation successfully, the expressionof green fluorescent protein could be observed by fluorescence microscopy.We can see the expression of Nanog gene is reduced by RT-PCR andWestern Blot, and select out the the most efficient recombinant plasmid.(2)CCK-8test showed the diminished capacity of cell proliferation, flowcytometry showed cell cycle arrested at the S phase, apoptosis cellsincreased, transwell tests showed invasion and migration capacitydiminished.Conclusion: After transfect by shRNA-Nanog, the proliferation ofgastric cancer cell line SGC-7901was significantly inhibited, cell cyclearrested at S phase and apoptosis cells increased, migration and invasioncapacity of cells became weaker. Above all, Nanog genes closely relatedwith cell cycle, apoptosis, proliferation, migration and invasion abilities ingastric cancer cell line SGC-7901.