ABSTRACT
OBJECTIVE：To investigate the effects and m echanism of oleanolic acid on inhibiting the proliferation ，invasion and metastasis of human ovarian cancer SKOV 3 cells. METHODS ：CCK-8 assay was used to detect the effects of different concentrations of oleanolic acid （10，20，40，60，80，100 μmol/L）on the proliferation of ovarian cancer SKOV 3 cells at 12，24， 36 and 48 h. The effects of low-dose and high-dose of oleanolic acid （20，40 μmol/L）on the metastasis and invasion ability of SKOV3 cells for 24 h were observed in Transwell assay. Western blotting assay was used to detect the effects of low-dose and high-dose of oleanolic acid on the protein expression of NF-κB p65，PRL-3，TNF-α，IL-6 and E-cadherin in SKOV 3 cells. Through LPS induction and NF-κB p65 plasmid transfection ，Western blotting and RT-qPCR assay were used to investigate the effects of low-dose and high-dose oleanolic acid on the expression of NF-κB/PRL-3 pathway related proteins and their mRNA. RESULTS ： With the increase of the concentration and action time of oleanolic acid ，the proliferation capacity of ovarian cancer SKOV 3 cells was decreased ，the surval rates of administration groups were significantly lower than that of the control group （P＜0.05 or P＜ 0.01）. Low-dose and high-dose of oleanolic acid could significantly reduce the number of migrating and invading cells （P＜0.05 or P＜0.01）. The protein relative expression of NF-κB p65，PRL-3，TNF-α and IL-6 in SKOV 3 cells were significantly decreased ， while the protein relative expression of E-cadherin was significantly increased （P＜0.05 or P＜0.01）. After LPS induction ，protein and mRNA relative expression of NF-κB p65，PRL-3，TNF-α and IL-6 were increased significantly in LPS model group ，while protein and mRNA relative expression of E-cadherin were significantly decreased （P＜0.05 or P＜0.01）. The protein and mRNA relative expression of NF-κ B p65，PRL-3，TNF-α and IL-6 were significantly decreased ，and protein and mRNA relative expression of E-cadherin were significantly increased in low-dose and high-dose of oleanolic acid group （P＜0.05 or P＜0.01）. In SKOV3 cells with over-expressed NF-κB p65，low-dose and high-dose of oleanolic acid c ould significantly down-regulat the proteinexpression of NF-κ B p65，PRL-3，TNF-α and IL-6，while upregult the protein relative expression of E-cadherin （P＜0.05 E-mail：122821905@qq.com or P＜0.01）. CONCLUSIONS ： Oleanolic acid can inhibit SKOV3 cells proliferation，invasion and metastasis by regulating NF-κB/PRL-3 signaling pathway.
ABSTRACT
@#To study the inhibitory effect of gigantol on proliferation, migration and invasion of human osteosarcoma U20S cells and to explore the mechanism. Methods: After being treated with different concentrations (10, 25, 50, 75, 100, 150 µmol/L) of gigantol for 24 and 48 h, the proliferation of U20S cells was detected by CCK-8 assay. Transwell assay was used to detect the effects of 25 µmol/L and 50 µmol/L gigantol on the migration and invasion abilities of U20S cells. The lipopolysaccharide (LPS) was used to induce inflammatory reaction in U20S cells before gigantol treatment; qPCR and WB were used to detect the mRNA and protein expressions of NF-κB (p65), TNF-α, IL-6 and PRL-3, respectively. Results: Different concentrations of gigantol could all inhibit the proliferation of sarcoma U20S cells at different time (P<0.05 or P<0.01). The 25 µmol/L and 50 µmol/L of gigantol could significantly inhibit the migration and invasion of osteosarcoma U20S cells (all P<0.01); at the same time, it could inhibit the protein expressions of NF-κB, TNF-α, IL-6 and PRL-3 (P<0.05 or P<0.01). After LPS induction, the mRNA and protein expressions of NF-κB, TNF-α, IL-6 and PRL-3 in U20S cells were significantly increased (all P<0.01); however, the consequent treatment with gigantol (25 and 50 µmol/L) reversed the effects of LPS on U20S cells obviously (P<0.05 or P<0.01). Conclusion: Gigantol can inhibit the proliferation, migration and invasion of osteosarcoma U20S cells, and its mechanism may be related to the regulation of NF-κB/PRL-3 signaling pathway.
ABSTRACT
The phosphatase of regenerating liver(PRL)families of phosphatases,consisting of PRL-1,PRL-2,and PRL-3,are individually overexpressed in a variety of cancer cell lines and tissues when eom-pared with their normal counterparts.Several recent studies have shown that PRL-3 is expressed at a higher lev-el and at a greater frequency in colorectal cancer with liver metastases compared with primary colorectal tumorsand normal colon tissue.Expression of PRL can enhance metastatic and invasive properties of cells and initiate tumor angiogenesis in experimental mice.However,the exact mechanism and in- teracting proteins of the PRL remain unclear.With further study,PRL-3 may server as an attractive target for therapeutic intervention and marker for colon cancer metastasis.
ABSTRACT
PURPOSE: Overexpression of the protein tyrosine phosphatase (PRL-3) is elevated in liver metastases derived from colorectal cancer. We examined PRL-3 expression in the primary lesion of colorectal cancer patients and investigated its relation to clinicopathological features. METHODS: A total of 63 randomly selected patients who underwent surgical resection for colorectal cancer between May 2001 and June 2005 at our hospital were investigated. Formalin-fixed and paraffin-embedded specimens from colorectal cancer patients who underwent surgical resections for primary tumors were collected. The expression of PRL-3 was detected by immunohistochemistry and the relation with age, sex, primary tumor size, tumor cell differentiation, depth of invasion, microscopic lymph node metastases, vascular invasion, numbers of lymph node metastases, postoperative stage, and postoperative survival time were analyzed. RESULTS: A total of 16 of the 63 colorectal cancer patients were detected with liver metastases during the follow-up periods. Liver resection was performed for those liver metastases patients. Five patients developed lung metastases after liver resection. PRL-3 expression was detected in 46 colorectal cancer patients. Fourteen patients with lymphatic invasion had positive expression of PRL-3 that was significant (P=0.042). The incidence of PRL-3 expression in the T stage was significant (P=0.019). Moreover, PRL-3 expression was closely associated with liver metastases (P=0.048). CONCLUSIONS: These results indicate that an investigation of PRL-3 expression in primary colorectal cancer lesions may contribute to the detection of occult liver metastases and to a differentiation between postoperative management strategies.
Subject(s)
Humans , Cell Differentiation , Colonic Neoplasms , Colorectal Neoplasms , Follow-Up Studies , Immunohistochemistry , Incidence , Liver , Lung , Lymph Nodes , Neoplasm Metastasis , Protein Tyrosine Phosphatases , Rectal NeoplasmsABSTRACT
PRL-3 is a key gene related to metastasis of colorectal carcinoma. However, it is known little about the possible regulatory mechanisms of PRL-3 gene expression. There were three possible promoter regions predicted by TRED, a promoter prediction software,which were all located in the upstream regions of PRL-3 gene. One of PRL-3 gene candidate promoters was located in the region of about -1kb upstream proximal to 5′ UTR of PRL-3 gene. Many possible transcription factor binding sites such as Snail, n-MYC, ARNT, E74A, NF-kappaB, NRF-2 and AML-1 were predicted in the region by Consite, a promoter analysis web system. Interestingly, a 5′ CACCTG 3′ core sequence and other related sequences of snail binding sites were found in promoter region of PRL-3 genes. Two PRL-3 gene promoters between -699 to 299 nt and between -642 to -383 nt were cloned into pGL3 vector with luciferase report gene. Both of them had promoter activities in four different cell lines including colorectal carcinoma cell lines SW480 and SW620, nasopharyngeal carcinoma cell line CNE2 and human embryo kidney cell line 293A. Interestingly, the luciferase activities of the short DNA fragmentations with Snail binding site′s core sequence 5′ CACCTG 3′ were higher than that of the longer one. PRL-3 promoter obtaining the 5′ CACCTG 3′ core sequence of Snail binding sites, was validated to bind to snail by chromatin immunoprecipitation (CHIP) analysis and electrophoretic mobility shift assay (EMSA) in SW480 cells. The data suggested that Snail was involved in regulation of PRL-3.