Teera Chewonarin

Momordica charantia leaf extract suppresses rat prostate cancer progression in vitro and in vivo

Cancer metastasis is a major cause of death in cancer patients, with invasion as a first step greatly contributing to the failure of clinical treatments. Any compounds with an inhibitory influence on this process are therefore of prime interest. Momordica charantia (bitter melon) is widely consumed as a vegetable and especially as a folk medicine in Asia. Here, we investigated the anti‐invasive effects of bitter melon leaf extract (BMLE) on a rat prostate cancer cell line (PLS10) in vitro and in vivo. The results indicated that non‐toxic concentrations of BMLE significantly inhibited the migration and invasion of cells in vitro. The results of zymography showed that BMLE inhibited the secretion of MMP‐2, MMP‐9 and urokinase plasminogen activator (uPA) from PLS10. Real‐time RT‐PCR revealed that BMLE not only significantly decreased gene expression of MMP‐2 and MMP‐9, but also markedly increased the mRNA level of TIMP‐2, known to have inhibitory effects on the activity of MMP‐2. An EnzChek gelatinase/collagenase assay showed that collagenase type IV activity was partially inhibited by BMLE. In the in vivo study, intravenous inoculation of PLS10 to nude mice resulted in a 100% survival rate in the mice given a BMLE‐diet as compared with 80% in the controls. The incidence of lung metastasis did not show any difference, but the percentage lung area occupied by metastatic lesions was slightly decreased in the 0.1% BMLE treatment group and significantly decreased with 1% BMLE treatment as compared with the control. Thus, the results indicate for the first time an anti‐metastatic effect of BMLE both in vitro and in vivo. (Cancer Sci 2010)

Silencing of connexin 43 suppresses invasion, migration and lung metastasis of rat hepatocellular carcinoma cells

To reduce cancer mortality, understanding of mechanisms of cancer metastasis is crucial. We have established six rat hepatocellular carcinoma (HCC) cell lines, which exhibit differing metastatic potential to the lung after inoculation into the tail veins of nude mice. In the present experiment, we investigated the process of cell attachment to metastatic sites and possible regulating factors. One hour after inoculation, two of two HCC cell lines with high metastatic potential and one of two HCC cell lines with low metastatic potential exhibited many attached cells in the lung. One day after inoculation, lung metastatic foci were observed only with highly‐metastatic cells with elevated connexin 43 (Cx43) expression as assessed by cDNA array analysis. Furthermore, 24 or 48 h after transfection of an siRNA targeting Cx43, in vitro invasion and migration were suppressed by 68% (P < 0.001) and 36% (P < 0.05) compared with control‐siRNA transfected cells, despite no differences in cellular morphology, cell proliferation or apoptotic activity. Moreover, the number of metastatic nodules per lung area in nude mice was significantly (P < 0.01) reduced. In conclusion, suppression of Cx43 expression in tumor cells reduced in vitro migration and invasion capacity and in vivo metastatic ability so that Cx43 has potential as a molecular target for prevention of cancer metastasis with Cx43 overexpressing tumors. (Cancer Sci 2012; 103: 860–867)

Particulate matter, PM 10 & PM 2.5 levels, and airborne mutagenicity in Chiang Mai, Thailand

Daily levels of particulate matter (PM) in the ambient air (PM 2.5 and PM 10) were measured in a northern city of Thailand (Chiang Mai) from March 1998 to October 1999. Twenty-four-hour air particulate matter samples were collected each day with Airmetric Minivol portable air samplers. Monthly averages of PM 2.5 from four stations in Chiang Mai varied from 15.39 to 138.31microg/m(3) and 27.29 to 173.40 microg/m(3) for PM 10. The PM 2.5 annual average was 58.48 mg/m(3) and PM 10, 86.38 microg/m(3). Daily PM 2.5 (24h values) during the winter months in Chiang Mai frequently exceeded 200-300 microg/m(3). The maximum concentrations of PM 2.5 (24h average) in Chiang Mai air from December 1998 to April 1999 were 2.8-, 3.5-, 4.2-, 6.5- and 3.2-fold higher than the US Environmental Protection Agency (US EPA), PM 2.5, 24h standard of 65 microg/m(3). From May to October, the mean 24h levels of PM 2.5 and PM 10 were at acceptable levels. The data shows that during the winter season (December to March), levels of PM 2.5 and PM 10 in the Chiang Mai atmosphere are very high, and there may be significant health implications associated with these high concentrations. During the summer season, the fine particles were generally within the acceptable levels. To our knowledge, these are the first measurements of PM 2.5 to be reported for the city of Chiang Mai and they indicate considerable ambient fine particle exposures to the Chiang Mai population. In addition, dichloromethane extracts of airborne particulate matter PM 2.5 or PM 10 collected in the months of winter in the city of Chiang Mai were mutagenic to Salmonella typhimurium strain TA100 without metabolic activation. The mutagenicity appeared to track particle concentrations and increased in the presence of S9 mix.

Ellagic Acid Inhibits Migration and Invasion by Prostate Cancer Cell Lines

Polyphenolic compounds from pomegranate fruit extracts (PFEs) have been reported to possess antiproliferative, pro-apoptotic, anti-inflammatory and anti-invasion effects in prostate and other cancers. However, the mechanisms responsible for the inhibition of cancer invasion remain to be clarified. In the present study, we investigated anti-invasive effects of ellagic acid (EA) in androgen-independent human (PC-3) and rat (PLS10) prostate cancer cell lines in vitro. The results indicated that non-toxic concentrations of EA significantly inhibited the motility and invasion of cells examined in migration and invasion assays. The EA treatment slightly decreased secretion of matrix metalloproteinase (MMP)-2 but not MMP-9 from both cell lines. We further found that EA significantly reduced proteolytic activity of collagenase/gelatinase secreted from the PLS-10 cell line. Collagenase IV activity was also concentration-dependently inhibited by EA. These results demonstrated that EA has an ability to inhibit invasive potential of prostate cancer cells through action on protease activity.

Effects of Nobiletin on PhIP-Induced Prostate and Colon Carcinogenesis in F344 Rats

The current study was designed to investigate the effects of nobiletin (5,6,7,8,3′,4′-hexamethoxy flavone) on 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP)-induced prostate and colon carcinogenesis. PhIP was administered to 6-wk-old F344 male rats intragastrically (100 mg/kg) twice a wk for 10 wk. The animals were given 0.05% nobiletin or the basal diet for 50 wk. At the end of the experiment, serum testosterone, estrogen, and leptin did not differ between the 2 groups. The body weights of nobiletin-treated rats were significantly higher than controls (P < 0.05), and feeding of nobiletin significantly reduced the relative prostate (P < 0.05) and testes (P < 0.05) weights as well as the Ki67 labeling index in the normal epithelium in the ventral prostate (P < 0.01). The incidence and multiplicity of adenocarcinomas in nobiletin-treated ventral prostate were 50% and 36%, respectively, of controls, but the differences were not statistically significant. However, nobiletin did significantly reduce the total number of colonic aberrant crypt foci (ACF) compared to the control value (P < 0.05). Nobiletin, therefore, may have potential for chemoprevention of early changes associated with carcinogenesis in both the prostate and colon.

Ellagic acid, a component of pomegranate fruit juice, suppresses androgen-dependent prostate carcinogenesis via induction of apoptosis

Ellagic acid (EA), a component of pomegranate fruit juice (PFJ), is a plant‐derived polyphenol and has antioxidant properties. PFJ and EA have been reported to suppress various cancers, including prostate cancer. However, their chemopreventive effects on development and progression of prostate cancer using in vivo models have not been established yet.

Expression of glutathione peroxidase 2 is associated with not only early hepatocarcinogenesis but also late stage metastasis.

Understanding of mechanisms of cancer progression is very important for reduction of cancer mortality. Of six rat hepatocellular carcinoma (HCC) cell lines, differing in their metastatic potential to the lung after inoculation into the tail vein of nude mice, the most metastatic featured particular overexpression of glutathione peroxidase 2 (GPX2). Therefore, we analyzed the influence of interference in highly metastatic L2 cells by siRNA transfection. Gpx2 siRNA significantly inhibited cell proliferation at 24 and 48h time points with induction of apoptosis but not cell cycle arrest. High expression of mutated p53 was detected in all HCC cell lines, with reduction in Gpx2 siRNA-transfected cells. Migration and invasion in vitro were also suppressed as compared to control siRNA-transfected cells and secretion of matrix metalloproteinase 9 was reduced. In vivo, the numbers and areas of metastatic nodules per area in the lungs were significantly reduced in the mice inoculated with Gpx2 siRNA-transfected cells as compared to control siRNA-transfected cells. In conclusion, expression of GPX2 is associated with cancer metastasis from rat HCCs both in vitro and in vivo. Together with immunohistochemical findings of elevated expression in rat and also human liver lesions, the results point to important roles in hepatocarcinogenesis.

Purple Rice Extract Supplemented Diet Reduces DMH- Induced Aberrant Crypt Foci in the Rat Colon by Inhibition of Bacterial β-Glucuronidase

BACKGROUND Purple rice has become a natural product of interest which is widely used for health promotion. This study investigated the preventive effect of purple rice extract (PRE) mixed diet on DMH initiation of colon carcinogenesis. MATERIALS AND METHODS Rats were fed with PRE mixed diet one week before injection of DMH (40 mg/kg of body weight once a week for 2 weeks). They were killed 12 hrs after a second DMH injection to measure the level of O6-methylguanine and xenobiotic metabolizing enzyme activities. RESULTS In rats that received PRE, guanine methylation was reduced in the colonic mucosa, but not in the liver, whereas PRE did not affect xenobiotic conjugation, with reference to glutathione-S-transferase or UDP-glucuronyl transferase. After 5 weeks, rats that received PRE with DMH injection had fewer ACF in the colon than those treated with DMH alone. Interestingly, a PRE mixed diet inhibited the activity of bacterial β-glucuronidase in rat feces, a critical enzyme for free methylazoxymethanol (MAM) release in the rat colon. These results indicated that purple rice extract inhibited β-glucuronidase activity in the colonic lumen, causing a reduction of MAM-induced colonic mucosa DNA methylation, leaded to decelerated formation of aberrant crypt foci in the rat colon. CONCLUSIONS The supplemented purple rice extract might thus prevent colon carcinogenesis by the alteration of the colonic environment, and thus could be further developed for neutraceutical products for colon cancer prevention.

Inhibitory Effects of Dried Longan (Euphoria longana Lam.) Seed Extract on Invasion and Matrix Metalloproteinases of Colon Cancer Cells

The critical step in colorectal cancer progression and associated mortality is cancer invasion, which depends on two key gelatinase enzymes, matrix metalloproteinases-2 and -9. Dried longan ( Euphoria longana Lam.) seed is a rich natural source of antioxidant polyphenols.This study evaluated the effect of dried longan seeds on colon cancer cell invasion via gelatinase function and expression. Three dried longan seed fractions were collected by Sephadex LH-20 column chromatography. They showed a potent inhibitor on colorectal cancer cell invasion and gelatinase activity. The antigelatinase activities of fractions 1 and 2 were a direct effect via Zn²⁺ chelation, whereas fraction 3 modulated indirectly through suppression of zymogen activators. Among the fractions, only fraction 3 reduced the gelatinase expression, which was correlated with the levels of tissue inhibitor of metalloproteinase-1 and may as well involve the p38 mitogen-activated protein kinases and the c-Jun N-terminal kinase signaling pathways. This primary research has manifested and encouraged the anticancer properties of dried longan seed extracts with potential inhibitory effects on cancer cell invasion as well as antigelatinase activity and expression in colon cancer cells.

Aflatoxin–albumin adduct formation after single and multiple doses of aflatoxin B1 in rats treated with Thai medicinal plants

The objective was to conduct an assessment of the ability of two Thai medicinal plants, Cymbopogon citratus Stapf and Murdannia loriformis, to modulate levels of serum aflatoxin-albumin (AF-albumin) adducts following aflatoxin B(1) (AFB(1)) exposure in rats. The influence of the plant extracts on AF-albumin adduct formation after a single exposure to 250 microg/kg body weight (bw) AFB(1) was measured over a 48-h period. Rats received M. loriformis extract (3 g/kg bw) or C. citratus Stapf extract (5 g/kg bw) daily for the week prior to the AFB(1) administration. In control rats, maximum adduct levels were observed 12 h post-AFB(1) treatment but in the animals receiving Murdannia extract, maximum levels occurred earlier, at 4 h post-treatment. No such effect was observed with the Cymbopogon extract. Daily treatment of rats with AFB(1) at 250 microg/kg bw for 3 weeks caused serum AF-albumin adduct levels to accumulate over a 10-14 day period and reach plateau levels 4.4-fold higher than observed after a single dose. Treatment with Murdannia extract for 1 week before and then throughout the AFB(1) exposure period resulted in a slight decrease in the AF-albumin adduct levels in the first week of the intervention. After that time, however, the reduction in adduct levels in the Murdannia extract group did not differ significantly from controls. No significant alteration in the biomarker levels was seen with the Cymbopogon extract treatments compared to control rats.