ABSTRACT
BACKGROUND: Vascular endothelial growth factor (VEGF) is involved in the growth of new blood vessels that feed tumors and kinesin spindle protein (KSP) plays a critical role in mitosis involving in cell proliferation. Simultaneous silencing of VEGF and KSP, an attractive and viable approach in cancer, leads on restricting cancer progression. The purpose of this study is to examine the therapeutic potential of dual gene targeted siRNA cocktail on human hepatocellular carcinoma Hep3B cells. RESULTS: The predesigned siRNAs could inhibit VEGF and KSP at mRNA level. siRNA cocktail showed a further downregulation on KSP mRNA and protein levels compared to KSP-siRNA or VEGF-siRNA, but not on VEGF expression. It also exhibited greater suppression on cell proliferation as well as cell migration or invasion capabilities and induction of apoptosis in Hep3B cells than single siRNA simultaneously. This could be explained by the significant downregulation of Cyclin D1, Bcl-2 and Survivin. However, no sigificant difference in the mRNA and protein levels of ANG2, involving inhibition of angiogenesis was found in HUVECs cultured with supernatant of Hep3B cells treated with siRNA cocktail, compared to that of VEGF-siRNA. CONCLUSION: Silencing of VEGF and KSP plays a key role in inhibiting cell proliferation, migration, invasion and inducing apoptosis of Hep3B cells. Simultaneous silencing of VEGF and KSP using siRNA cocktail yields promising results for eradicating hepatocellular carcinoma cells, a new direction for liver cancer treatment.
Subject(s)
Humans , Kinesins/genetics , Apoptosis/genetics , Gene Silencing , RNA, Small Interfering/genetics , Vascular Endothelial Growth Factor A/genetics , Cell Proliferation/genetics , Tetrazolium Salts , Transfection , Cysteine Proteinase Inhibitors/metabolism , Down-Regulation , Cell Movement , Blotting, Western , Kinesins/metabolism , Annexin A5 , Genes, bcl-2 , Cyclin D1/metabolism , Vesicular Transport Proteins/metabolism , Cell Line, Tumor , Vascular Endothelial Growth Factor A/metabolism , Inhibitor of Apoptosis Proteins/metabolism , Human Umbilical Vein Endothelial Cells/metabolism , Real-Time Polymerase Chain Reaction , Flow Cytometry , Survivin , Mitosis/geneticsABSTRACT
OBJECTIVE: Endometriosis is the presence of normal endometrial mucosa (glands and stroma) abnormally implanted on the outside of uterus. The pathophysiology of endometriosis is not clear yet, but Sampson's theory of the transplantation of endometrial tissue onto the pelvic peritoneum via retrograde menstruation is most widely accepted. Vascular endothelial cell growth factor (VEGF) is involved in the pathophysiology of endometriosis via its angiogenetic property. This study was designed to investigate whether single nucleotide polymorphism and its haplotype and diplotype of VEGF genes are associated with the risk of advanced endometriosis or not. METHODS: This study investigated 260 patients of endometriosis; they underwent operation, and were diagnosed with endometriosis stage III, IV histopathologically. And control group of 199 women underwent surgery with benign ovarian cyst. The single nucleotide polymorphisms of VEGF gene were -2578C>A, 405G>C, 936C>T. They were analyzed by polymerase chain reaction and restriction fragment length polymorphism, and haplotype and diplotype analysis were done. RESULTS: The result of this study showed no association among -2578C/A, 405G>C, 936C>T single nucleotide polymorphisms and severe endometriosis. Haplotype and diplotype of -2578C>A, 405G>C, 936C>T in the VEGF gene were shown to have no association with endometriosis. We found no association between VEGF genetic polymorphism and risk of endometriosis. And haplotype and diplotype analysis also revealed no statistically significant value between VEGF polymorphism and endometriosis. CONCLUSIONS: So, the results of this study suggest polymorphism of VEGF gene may not be associated with risk of endometriosis in Korean women.
Subject(s)
Female , Humans , Endometriosis , Endothelial Cells , Haplotypes , Menstruation Disturbances , Mucous Membrane , Ovarian Cysts , Peritoneum , Polymerase Chain Reaction , Polymorphism, Genetic , Polymorphism, Restriction Fragment Length , Polymorphism, Single Nucleotide , Transplants , Uterus , Vascular Endothelial Growth Factor AABSTRACT
OBJECTIVE:To explore the inhibitory effects and mechanism of Rhizoma Sparganii and Rhizoma Curcumae on the angiogenesis of new granulation tissue in sponge.METHODS:The experimental endometriosis in female rats was established with sponge implantation.The effects of Rhizoma Sparganii and Rhizoma Curcumae on the angiogenesis of new granulation tissue in sponge were observed after intragastric medication for 2 consecutive weeks.The staining of endothelial cell specific Ⅷ factor and vascular endothelial cell growth factor(VEGF) was carried out by immunohistochemistry method.The expression of VEGF mRNA was detected by RT-PCR assay.RESULTS:In rats receiving high dosage of Rhizoma Sparganii and Rhizoma Curcumae(SC),the area of new granulation tissue in sponge,the number of blood vessels,the expression of VE-GF and VEGF mRNA were all significantly lower than in model group(P
ABSTRACT
Objective:To study the relation between clinical index and prognosis and the level of vascular endothelial cell growth factor(VEGF) in serum of non small cell lung cancer(NSCLC) patients before and after radiotherapy.Methods:VEGF serum of 39 patients with NSCLC and in 20 healthy individuals as control were analyzed with double sandwich in assay ELISA.Results:The level of VEGF serum elevated significantly before radiotherapy in NSCLC patients group as compared with the control(P0 05).And the level gradually declined 0,2～3 months after radiotherapy (P