ABSTRACT
BACKGROUND: Human gallbladder cancer (GBC) is an aggressive malignant neoplasm with a poor prognosis. The development of ideal tools for example tumor cell lines for investigating biological behavior, metastatic mechanism and potential treatment in GBCs is essential. In present study, we established and characterized a GBC cell line derived from primary tumor. METHODS: Primary culture method was used to establish this cell line from a primary GBC. Light and electron microscopes, flow cytometry, chromosome analysis, heterotransplantation and immunohistochemistry were used to characterize the epidemic tumor characteristics and phenotypes of this cell line. RESULTS: A novel GBC cell line, named TJ-GBC2, was successfully established from primary GBC. This cell line had characteristic epithelial tumor morphology and phenotypes in consistent with primary GBC, such as polygon and irregular cell shape, increased CA19-9 and AFP levels, and positive expression of CK7, CK8, CK19 and E-cadherin with negative vimentin. Moreover, about 25% of the cells were in the S-G2/M phase; abnormity in structure and number of chromosome with a peak number of 90-105 and 80% hypertetraploid were observed. Furthermore, this cell line had higher invasion and highest migration abilities compared to other GBC cell lines; and metastatic-related marker MMP9 and nm23 were positively expressed. CONCLUSIONS: A novel highly aggressive GBC cell line TJ-GBC2 was successfully established from primary GBC. TJ-GBC2 cell line may be efficient tool for further investigating the biological behaviors, metastatic mechanism and potential targeted therapy of human GBC.
ABSTRACT
AIM: To study the effect of gum mastic, a natural resin, on the proliferation of androgen-independent prostate cancer PC-3 cells, and further investigate the mechanisms involved in this regulatory system, taking nuclear factor kappaB (NF-kappaB) signal as the target. METHODS: 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and a flow cytometer were used to detect the effect of gum mastic on the proliferation of PC-3 cells. Then, reporter gene assay, RT-PCR, and Western blotting were carried out to study the effects of gum mastic on the NF-kappaB protein level and the NF-kappaB signal pathway. The expression of genes involved in the NF-kappaB signal pathway, including cyclin D1, inhibitors of kappaBs (I kappaB alpha), and phosphorylated Akt (p-AKT), were measured. In addition, transient transfection assays with the 5X NF-kappaB consensus sequence promoter was also used to test the effects of gum mastic. RESULTS: Gum mastic inhibited PC-3 cell growth and blocked the PC-3 cell cycle in the G1 phase. Gum mastic also suppressed NF-kappaB activity in the PC-3 cells. The expression of cyclin D1, a crucial cell cycle regulator and an NF-kappaB downstream target gene, was reduced as well. Moreover, gum mastic decreased the p-AKT protein level and increased the I kappa B alpha protein level. CONCLUSION: Gum mastic inhibited the proliferation and blocked the cell cycle progression in PC-3 cells by suppressing NF-kappaB activity and the NF-kappaB signal pathway.
Subject(s)
Antineoplastic Agents, Phytogenic/pharmacology , NF-kappa B/drug effects , Prostatic Neoplasms/drug therapy , Resins, Plant/pharmacology , Androgens/physiology , Cell Nucleus/chemistry , Cell Nucleus/metabolism , Cell Proliferation/drug effects , Cyclin D1/biosynthesis , Cyclin D1/genetics , Humans , Male , Mastic Resin , Oncogene Protein v-akt/biosynthesis , Oncogene Protein v-akt/genetics , Reverse Transcriptase Polymerase Chain Reaction , Signal Transduction/drug effectsABSTRACT
OBJECTIVE: To study the effects of low-power electromagnetic fields of different frequencies on proliferation and DNA damage of gallbladder cancer cells. METHODS: The cell growth curve was drawn and single cell gel electrophoresis performed to evaluate the proliferation and DNA damage of gallbladder cancer cells respectively after the cells were exposed to electromagnetic fields of different frequencies. RESULTS: After exposure to low-power electromagnetic fields of different frequencies (0.1-40 MHz), the cells displayed significant changes with obvious cell proliferation inhibition and DNA strand breakage. CONCLUSION: Low-power electromagnetic fields within the range of 0.1-40 MHz may impair the DNA strand and cause inhibition of proliferation of the gallbladder cancer cells, and these effects are related to the frequency of the electromagnetic fields but not in a linear fashion.