RESUMO
AIM: To explore the effect of luteolin on the metastasis of ovarian carcinoma HO-8910PM cells in vitro, and to elucidate its mechanisms. METHODS: The in vitro invasion and mobility of HO-8910PM were measured by Boyden chamber assay after exposure to luteolin for 6 h. Gelatinase activity was tested by gelatin zymography assay. The expression of tissue inhibitor of metalloproteinase-1 (TIMP-1), nm23 mRNA and that of ERK2 protein were tested by RT-PCR and Western blotting, respectively. RESULTS: Luteolin significantly inhibited in vitro invasiveness and mobility in HO-8910PM cells in a dose dependent manner as measured by Boyden chamber assay. Luteolin inhibited the MMP-9 secretion from HO-8910PM cells. Luteolin did not affect the mRNA expression of TIMP-1 and nm23. Luteolin also decreased ERK2 expression. CONCLUSION: Luteolin dose-dependently inhibited the metastasis of HO-8910PM cells in vitro. Inhibition of MMP-9 secretion and ERK2 expression may be involved in the anti-metastasic process of luteolin.
RESUMO
AIM: Open reading frame(ORF) of death associa ted protein kinase1(DAPK1) gene was cloned for studying on tumor forming and met astasis.METHODS: Based on nucleotide sequence of DAPK1 gene f rom GenBank, a pair of primers was designed. DAPK1 gene ORF was transfected into Raji cells in expression vector pcDNA3.1(+) with lipofectamine reagent. Morphol ogic assessment of apoptosis was performed with fluorescence microscope cytotoxi city and cell viability was assayed by MTT. RESULTS: DAPK1 gene ORF was amprified from K562 cells by RT-PCR. It was cloned into plasmid pMD18-T and sequenced. There were seven mutation in 4 300 bp nucleotide sequence rel ativel y to DAPK1 nucleotide sequence from GenBank, but six was synonymous mutation and one was single nucleotide polymorphism. 4 300 bp nucleotide of DAPK1 gene O RF was transfected into Raji cells. DAPK1 gene expression was detected in 48 h a fter it was transfected into Raji cells. Then Raji cells showed apoptosis.CONCLUS ION: Large fragment gene was cloned by RT-PCR and transfected into Raji cells successfully. Over-expression of DAPK1 gene induced Raji cells apoptosis.
RESUMO
Aim To investigate the inhibitory effect and its posssible mechanism of 5F from Pteris semipinnata L on the metastasis-associated ability of human highly metastatic ovarian carcinoma HO-8910PM cells in vitro.Methods MTT assay was used to examine the effect of 5F on proliferation of HO-8910PM cells after 24 hours treatment; Transwell Chamber assay was performed to determine the effect of 5F on invasion and migratory capacity of the cells; Effect on adhesion potential of HO-8910PM cells was tested by cell-Matrigel adhesion assay; The expression levels of NF-?B(P65) and VEGF proteins were assessed by Western blot analysis.Results 5F significantly inhibited invasion, migration, and adhesion capacity of HO-8910PM cells in vitro, their inhibitory rates after treatment with 50 ?mol?L -1 5F for 6 h were (37.57?0.62)%, (28.42?0.67)%, (46.07?4.49)%. 5F d istinctly down-regulated the expressions of NF-?B(P65) and VEGF protein. Conclusion 5F can inhibit the migration, invasion and adhesion of HO-8910PM cells in vitro,Its possible mechanism may be involved in the reduction of the expression level of NF-?B(P65) and VEGF. 5F might be potential drugs to inhibit tumor metastasis.
