Rna Silencing Of Hyaluronidase Hyal1 Gene On Malignant Potential Of Human Breast Cancer Cell

PART ONE EXPERIMENTAL STUDY OF THE EFFECTS OF SIRNA-HYAL1 ON EXPRESSION OF THE HYALURONIDASE GENE HYAL1 IN HUMAN BREAST CANCER CELLS Objective: To study the effect of SiRNA-HYAL1 on the expression ofhyaluronidase gene in human breast cancer cells. Methods: Reverse transcription-polymerse chain reaction was applied Result: After transfection SiRNA-HYAL1, the expression levels ofHYAL1 were inhibited significantly in human breast cancer cell linesMDA-MB-231, MDA-MB-435S, ZR-75 and ZR-75-30(p<0.05). Conclusion: The expression of HYAL1 mRNA can be inhibited bytransfection of SiRNA-HYAL1 in human breast cancer cells.human breast cancer cells; SiRNA, RNA interference;HYAL1; RT-PCR;gene expression level PART TWO EXPERIMENTAL STUDY OF THE EFFECTS OFSIRNA-HYAL1 ON PROLIFERATION AND CELL CYCLEIN HUMAN BREAST CANCER CELLS BY INTERFERING THE HYALURONIDASE GENE HYAL1 Objective: To study the effects of SiRNA-HYAL1 on the growth,proliferation and cell cycle of human breast cancer cells. Methods: Cell culture technique, RNAi technique, MTT-assay andflow cytometric analysis were applied. Result: After transfection SiRNA-HYAL1, proliferation was inhibitedobviously in human breast cancer cell lines with different levels ofhyaluronidase, such as MDA-MB-231, MDA-MB-435S, ZR-75 andZR-75-30 by MTT-assay(p<0.05). Flow cytometry showed that cells in Sphase were reduced(p<0.05), with most other cells were arresting in G0/ G1phase of cell cycle(p<0.05). Conclusion: The growth and proliferation of human breast cancer cellscan be inhibited obviously by transfection SiRNA-HYAL1. PART THREE EXPERIMENTAL STUDY OF THE EFFECTS OF SIRNA-HYAL1 ON HUMAN BREAST CANCER CELLS INVASION POTENTIAL Objective: To study the effect of SiRNA-HYAL1 on human breastcancer cells invasion potential. Methods: In vitor experiments were carried out to determinepenetrating matrigel capacity of human breast cancer cells lines whichexpress different levels of hyaluronidase. Result: After transfection of SiRNA-HYAL1, the invasive capacitywas decreased in those human breast cancer cell lines with different levelsof hyaluronidase, such as MDA-MB-231, MDA-MB-435S, ZR-75 andZR-75-30 (p<0.01). Conclusion: By RNA interfering HYAL1, the invasive potential ofhuman breast cancer cells can be inhibited obviously. PART FOUR EXPERIMENTAL STUDY OF THE EFFECTS OF SIRNA-HYAL1 ON TUMOR ANGIOGENESIS Objective: To study the effects of SiRNA-HYAL1 on angiogenesis ofhuman breast cancer cells. Methods: A technique of co-culture of human breast cancer cells andendothelial cells of human umbilical vein (ECV-304) were applied.observed the morphology change of ECV-304 induced by human breastcancer cells expressing levels of hyaluronidase after RNA interferingHYAL1. Result: After SiRNA interfering HYAL1, the capacity of tubeformation of ECV-304 induced by human breast cancer cell linesexpressing levels of hyaluronidase, such as MDA-MB-231,MDA-MB-435S, ZR-75 and ZR-75-30 was inhibited obviously (p<0.01). Conclusion: By SiRNA interfering HYAL1, the capacity ofangiogenesis induced by human breast cancer cells inhibited obviously.