Hui-Ling Chiou

Cyanidin 3-Glucoside and Peonidin 3-Glucoside Inhibit Tumor Cell Growth and Induce Apoptosis In Vitro and Suppress Tumor Growth In Vivo

Abstract: Dietary polyphenols, including anthocyanins, are suggested to be involved in the protective effects of fruits and vegetables against cancer. However, anticancer effects of peonidin 3-glucoside have not been clearly demonstrated, with only limited studies being available concerning the inhibitory effect of cyanidin 3-glucoside for tumor cell growth. Therefore, in this study, we have isolated and identified the two bioactive compounds, peonidin 3-glucoside and cyanidin 3-glucoside, from Oryza sativa L. indica, to treat various cancer cells. The results showed that, among analyzed cell lines, HS578T was the most sensitive to peonidin 3-glucoside and cyanidin 3-glucoside. Treatment with peonidin 3-glucoside or cyanidin 3-glucoside resulted in a strong inhibitory effect on cell growth via G2/M arrest. Regarding cell cycle–related proteins, peonidin 3-glucoside treatment resulted in down-regulation of protein levels of cyclin-dependent kinase (CDK)-1, CDK-2, cyclin B1, and cyclin E, whereas cyanidin 3-glucoside could decrease the protein levels of CDK-1, CDK-2, cyclin B1, and cyclin D1. In addition, cyanidin 3-glucoside or peonidin 3-glucoside also induced caspase-3 activation, chromatin condensation, and cell death. Furthermore, anthocyanins from O. sativa L. indica were evidenced by their inhibition on the growth of Lewis lung carcinoma cells in vivo.

Silibinin inhibits the invasion of human lung cancer cells via decreased productions of urokinase-plasminogen activator and matrix metalloproteinase-2†

Cancer metastasis, involving multiple processes and various cytophysiological changes, is a primary cause of cancer death and may complicate the clinical management, even lead to death. Silibinin is a flavonoid antioxidant and wildly used for its antihepatotoxic properties and recent studies have revealed pleiotropic anticancer and antiproliferative capabilities of silibinin. In this study, we first observed that silibinin exerted a dose‐ and time‐dependent inhibitory effect on the invasion and motility, but hardly on the adhesion, of highly metastatic A549 cells in the absence of cytotoxicity. To look at the precise involvement of silibinin in cancer metastasis, A549 cells were treated with silibinin at various concentrations, up to 100 μM, for a defined period and then subjected to gelatin zymography, casein zymography and Western blot to investigate the impacts of silibinin on metalloproteinase‐2 (MMP‐2), urokinase plasminogen activator (u‐PA), and tissue inhibitor of metalloproteinase‐2 (TIMP‐2), respectively. The results showed that a silibinin treatment may decrease the expressions of MMP‐2 and u‐PA in a concentration‐ and time‐dependent manner and enhance the expression of TIMP‐2. Further analysis with semi‐quantitative RT‐PCR showed that silibinin may regulate the expressions of MMP‐2 and u‐PA on the transcriptional level while on the translational or post‐translational level for TIMP‐2.

Silibinin Inhibits Invasion of Oral Cancer Cells by Suppressing the MAPK Pathway

Oral squamous cell carcinoma (OSCC) is the most common malignancy of the oral cavity. Here, we provide molecular evidence associated with the anti-metastatic effect of silibinin by showing a marked inhibition of the invasion and motility of SCC-4 tongue cancer cells, with 89% and 66.4% of inhibition, respectively, by 100 μM of silibinin. This effect was associated with a reduced expression of MMP-2 and u-PA, together with an enhanced expression of TIMP-2 and PAI-1. Silibinin also exerted an inhibitory effect on the phosphorylation of ERK1/2. Additionally, pre-treatment of SCC-4 cancer cells with 10 and 20 μM of U0126, a specific MEK inhibitor, resulted in a reduced expression of MMP-2 (18.7 and 51.4%) and u-PA (19.2 and 48.9%) concomitantly with a marked inhibition of cell invasion (13.7 and 45.7%). Finally, silibinin was evidenced by its inhibition of the metastasis of Lewis lung carcinoma (LLC) cells in vivo. These results suggested that silibinin can reduce the invasion and metastasis of tumor cells, and such a characteristic may be of great value in the development of a potential cancer therapy.

The aberrant expression of cytosolic carbonic anhydrase and its clinical significance in human non-small cell lung cancer

This study was designed to elucidate the possible relationship between the expression of cytosolic carbonic anhydrase (CA) and non-small cell lung cancer (NSCLC). The activity and protein expression patterns of carbonic anhydrase I (CAI) and II (CAII) of 70 NSCLC patients were analyzed by CA activity analysis, immunoblotting and immunohistochemical staining. The results showed that the CA activity and protein expression were significantly decreased in both squamous cell carcinoma (SCC) and adenocarcinoma (AD) (P<0.001 and P<0.001). From our study, it was suggested that the reduction of CAI and CAII in both SCC and AD patients may promote tumor cell motility and contribute to tumor growth and metastasis.

Glabridin inhibits migration and invasion by transcriptional inhibition of matrix metalloproteinase 9 through modulation of NF-κB and AP-1 activity in human liver cancer cells

High mortality and morbidity rates for hepatocellular carcinoma in Taiwan primarily result from uncontrolled tumour metastasis. Glabridin, a prenylated isoflavonoid of licorice (Glycyrrhiza glabra) roots, is associated with a wide range of biological properties, such as regulation of energy metabolism, oestrogenic, neuroprotective, anti‐osteoporotic and skin whitening. However, the effect of glabridin on the metastasis of tumour cells has not been clarified.

MicroRNA gene polymorphisms and environmental factors increase patient susceptibility to hepatocellular carcinoma.

Background Micro RNAs (miRNAs) are small RNA fragments that naturally exist in the human body. Through various physiological mechanisms, miRNAs can generate different functions for regulating RNA protein levels and balancing abnormalities. Abnormal miRNA expression has been reported to be highly related to several diseases and cancers. Single-nucleotide polymorphisms (SNPs) in miRNAs have been reported to increase patient susceptibility and affect patient prognosis and survival. We adopted a case-control research design to verify the relationship between miRNAs and hepatocellular carcinoma. Methodology/Principal Findings A total of 525 subjects, including 377 controls and 188 hepatocellular carcinoma patients, were selected. Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and real-time PCR were used to analyze miRNA146a (rs2910164), miRNA149 (rs2292832), miRNA196 (rs11614913), and miRNA499 (rs3746444) genetic polymorphisms between the control group and the case group. The results indicate that people who carry the rs3746444 CT or CC genotypes may have a significantly increased susceptibility to hepatocellular carcinoma (adjusted odds ratio [AOR] = 2.84, 95% confidence interval [CI] = 1.88–4.30). In addition, when combined with environmental risk factors, such as smoking and alcohol consumption, interaction effects were observed between gene polymorphisms and environmental factors (odds ratio [OR] = 4.69, 95% CI = 2.52–8.70; AOR = 3.38, 95% CI = 1.68–6.80). Conclusions These results suggest that a significant association exists between miRNA499 SNPs and hepatocellular carcinoma. Gene-environment interactions of miRNA499 polymorphisms, smoking, and alcohol consumption might alter hepatocellular carcinoma susceptibility.

CCR2-64I gene polymorphism increase susceptibility to oral cancer

The purpose of this study was to investigate the impact of MCP-1 and its receptor CCR2 gene polymorphisms on the susceptibility and clinicopathological characteristics of oral cancer, as well as the synergistic effect between these gene polymorphisms and well-known risk factors including alcohol, tobacco, and areca consumptions. Using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) technique for polymorphism analysis, 344 healthy controls and 216 oral cancer patients were recruited to reveal a significant association between V64I CCR2 gene polymorphism and oral cancer susceptibility. After adjusting for other confounders, individuals with GA (AOR=1.84; 95%CI=1.10-3.20) or at least one A allele (AOR=1.78; 95%CI=1.05-3.02) had a higher risk for oral cancer, compared to GG genotypes. Moreover, results also revealed that for subjects with GA or at least one A allele of V64I CCR2 gene polymorphism, those exposed to environmental risk factors possessed a significantly higher risk for oral cancer than those unexposed subjects. Therefore, genetic polymorphism of CCR2-64I may contribute to the susceptibility to oral cancer.

Effect of hemodialysis on the plasma level of type IV collagenases and their inhibitors

OBJECTIVES Increased cell expression of matrix metalloproteinase-9 (MMP-9) was associated with the development of atherosclerosis and osseoarticular tissue destruction in hemodialysis patients. In this study, the pre- and post-HD plasma concentrations of type IV collagenases and their inhibitors in HD patients were examined. DESIGN AND METHODS Commercial ELISA kits and Zymography techniques were used to assay the parameters in 40 patients pre- and post-HD session. RESULTS After the hemodialysis process, MMP-9, MMP-2 and TIMP-2 levels were 124 +/- 72, 706 +/- 242 and 248 +/- 90 ng/mL, significantly different from the pre-HD values (187 +/- 148, 759 +/- 304 and 43 +/- 14 ng/mL). TIMP-1 were not affected by HD. Female subjects and patients with chronic glomerulonephritis had higher TIMP-2 than their counterparts (p < 0.05). The effect of gender on MMP-2 levels was interacted with that of membrane types (p < 0.01). CONCLUSION These results indicated that the hemodialysis process tends to decrease the overall activity of the peripheral plasma MMP system in HD patients.

Pterostilbene suppresses oral cancer cell invasion by inhibiting MMP-2 expression

Objective: Polyphenol compounds, present in a wide variety of natural plants, exhibit antioxidant and free radical scavenging ability and induce apoptosis in various cancer cells. However, the effect of pterostilbene on oral cancer cell metastasis has not been clarified. Research design and methods: The present study aimed to examine the anti-metastatic properties of pterostilbene in human oral squamous cell carcinoma (SCC)-9 cells. Results: In this study, pterostilbene treatment significantly inhibited migration/invasion capacities of SCC-9 cells in vitro. The results of zymography and western blotting revealed that the activities and protein levels of the MMP-2 and urokinase-type plasminogen activator (u-PA) was inhibited by pterostilbene. Western blot analysis also showed that pterostilbene inhibits the phosphorylation of Akt, extracellular signal-regulated kinase 1/2 and p38. Determinations of the mRNA levels, real-time polymerase chain reaction and promoter assays were conducted to evaluate the inhibitory effects of pterostilbene on MMP-2 and u-PA expression in SCC-9 cells. Such inhibitory effects were associated with the upregulation of tissue inhibitor of metalloproteinase-2, plasminogen activator inhibitor-1 and the downregulation of the transcription factors of NF-κB, SP-1 and CREB signaling pathways. Conclusions: Pterostilbene may have potential use as a chemopreventive agent against oral cancer metastasis.

Psoralen reverses docetaxel-induced multidrug resistance in A549/D16 human lung cancer cells lines

Chemotherapy is the recommended treatment for advanced-stage cancers. However, the emergence of multidrug resistance (MDR), the ability of cancer cells to become simultaneously resistant to different drugs, limits the efficacy of chemotherapy. Previous studies have shown that herbal medicine or natural food may be feasible for various cancers as potent chemopreventive drug. This study aims to explore the capablility of reversing the multidrug resistance of docetaxel (DOC)-resistant A549 cells (A549/D16) of psoralen and the underlying mechanisms. In this study, results showed that the cell viability of A549/D16 subline is decreased when treated with psoralen plus DOC, while psoralen has no effect on the cell proliferation on A549 and A549/D16 cells. Furthermore, mRNA and proteins levels of ABCB1 were decreased in the presence of psoralen, while decreased ABCB1 activity was also revealed by flow cytometry. Based on these results, we believe that psoralen may be feasible for reversing the multidrug resistance by inhibiting ABCB1 gene and protein expression. Such inhibition will lead to a decrease in ABCB1 activity and anti-cancer drug efflux, which eventually result in drug resistance reversal and therefore, sensitizing drug-resistant cells to death in combination with chemotherapeutic drugs.