Suppression of cancer metastasis
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[0157]Cell Lines and Culture.
[0158]SK-OV-3 cells (human ovarian carcinoma cell line; ATCC HTB-77) and SK-OV-3ip.1 cells, and CaSki cells (human cervical carcinoma cell line; ATCC CRL-1550) were maintained at 37° C. in McCoy's 5A and RPMI-1640 medium, respectively. MDA-MB-231 cells (human mammary adenocarcinoma cell line; ATCC HTB-26) and TS / A cells (murine mammary adenocarcinoma cell line) were maintained at 37° C. in Dulbecco's modified Eagle's medium (DMEM) / F12 medium and DMEM, respectively. PC-3 cells (human prostate adenocarcinoma cell line; ATCC CRL-1435) and 22Rv1 cells (human prostate carcinoma cell line; ATCC CRL-2505) were maintained at 37° C. in RPMI-1640 medium. All medium were supplemented with 10% fetal bovine serum (FBS), 2 mM L-glutamine, 100 units / ml penicillin, and 100 μg / ml streptomycin.
[0159]Cytotoxicity Assay.
[0160]Cell survival was determined by MTT assay or WST-1 assay. In brief, 1×105 cells / well of tumor cells were seeded in 96-well plates in serum-free...
example 2
rVP1 Suppressed Tumor Cell Metastasis In Vivo
[0181]A. In Vitro rVP1-Treated MDA-MB-231 Cells Followed by i.v. Injection into BALB / C Mice.
[0182]We then examined whether pretreated MDA-MB-231 human mammary adenocarcinoma cells with rVP1 for 24 h to inhibit metastatic ability in vitro then injected the cells intravenously in mice could reduce their metastasis in mice lung as compare with MDA-MB-231 cells without rVP1 pretreatment. Data showed that 0.1 and 0.2 μM rVP1-treated MDA-MB-231 cells significantly decreased cancer cells metastasis in mice lung tissue (FIG. 4A). The related lung weight and the number of tumor foci in lung were also significantly decrease compared to tumor group without rVP1 treatment (FIGS. 4B-C).
[0183]B. Orthotropic PC-3 Xenograft Model.
[0184]We also examined whether rVP1 could suppress prostate cancer metastasis in vivo. PC-3 human prostate adenocarcinoma cells were first orthotopically implanted to nude mice' prostate and 4-wks or 10-wks after implantation, r...
example 3
F-HSA Suppressed Tumor Cell Invasion and / or Migration in MDA-MB-231 Cells, PC-3 Cells, 22Rv1 Cells and CaSki Cells In Vitro
[0187]To examine whether other fibrillar proteins such as F-HSA also suppressed cancer cell invasion and / or migration, the effect of F-HSA on cancer cell invasion and / or migration were measured by using Boyden chamber assay. After being treated with various concentrations of F-HSA (0.1 μM to 0.2 μM F-HSA in MDA-MB-231 cells; 0.025 to 0.1 μM F-HSA in PC-3 cells; 0.025 to 0.05μM F-HSA in 22Rv1 cells; 0.2 to 0.4 μM F-HSA in CaSki cells), the invasion and / or migration abilities of a variety of cancer cells were significantly suppressed. It is of note that at these concentrations, F-HSA did not affect cell viability by using MTT assay (FIG. 7).
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