Effect And Mechanism Study Of PLD2 Gene Silencing On Prolifera Tion And Migration Of Colon Cancer Cells

Part one EXPRESSION AND SIGNIFICANCE OF PLD2 IN COLON CARCINOMAObjective: To measure the expression of PLD2 in colon cancer tissues and corresponding normal colon tissues, explore its relationship with clinical pathological features.Methods: 1. The protein expression of PLD2 in thirty colon adenocarcinoma specimens and their corresponding adjacent normal colonic mucosa tissues were measured by immunohistochemistry and the relationship between PLD2 expression and clinical pathological characteristics were analyzed by statistical software.2. The m RNA expression of PLD2 in colon cancer tissue and normal colon tissue were measured by quantitative real-time polymerase chain reaction(q RT-PCR).3. The total protein expression of PLD2 in colon carcinoma tissue,corresponding noncancerous colon mucosa tissue and different colon cancer cell lines.Results: 1. PLD2 is mainly expressed in the cytoplasm and cytomembrane of colon cancer cell, the PLD2 protein was expressed predominantly in colon cancer tissues(P<0.05), and was positively associated with the tumor size of colon cancer(P<0.05).2.The m RNA expression of PLD2 in colon cancer tissue was greatly elevated compared with normal colon tissue(P<0.05).3. The total protein of PLD2 was highly expressed in colon cancer tissue,SW480 and SW620 colon cancer cell lines(P<0.05).Conclusion:PLD2 was overexpressed in colon cancer tissue, SW480 and SW620 cancer cell lines, elevated expression of PLD2 was positively correlated with the size of colon cancer.Part two EFFECT OF PLD2 GENE SILENCING ON PROLIFERATION AND MIGRATION OF COLON CANCER CELLSObjective: To investigate the effect of silencing the PLD2(phospholipases D2) gene expression on cell proliferation and migration of colon cancer cell lines and to explore the underline molecular mechanisms.Methods: 1. The small interfering RNA(si RNA) targeted PLD2 was transfected into SW480 and SW620 cells by adenovirus. Expression level of PLD2 m RNA and protein were measured by quantitative real-time polymerase chain reaction(q RT-PCR) and Western blot, respectively.2. Plate clone formation assay was used to measure the proliferation of colon cancer cells.3. The migration of the cells were detected by transwell chamber assay.4.The expression of Cyclin D1 and Metalloproteinase-2(MMP-2) protein were measured by Western blot in both cell lines.Result:1. PLD2 si RNA greatly inhibited the expression of PLD2 in colon cancer cells at both m RNA and protein levels(all P<0.01).2. PLD2 gene silencing greatly inhibit the proliferation of both colon cancer cells(P=0.000).3. Down-regulated the PLD2 expression significantly suppressed the proliferation and migration ability of the cells(P=0.000).4. The expression of Cyclin D1 and MMP-2 proteins were both inhibited by the silencing of PLD2 gene.(P=0.000)Conclusion:Down-regulated PLD2 gene expression clearly inhibited the proliferation and migration of colon cancer cells may be closely associated with the decreased expression of Cyclin D1 and MMP-2, indicating a way of gene therapy for colon cancer.