Thymoquinone suppresses migration of Lo Vo human colon cancer cells by reducing prostaglandin E2 induced COX-2 activation

AIM To identify potential anti-cancer constituents in natural extracts that inhibit cancer cell growth and migration.METHODS Our experiments used high dose thymoquinone(TQ) as an inhibitor to arrest Lo Vo(a human colon adenocarcinoma cell line) cancer cell growth, which was detected by cell proliferation assay and immunoblotting assay. Low dose TQ did not significantly reduce Lo Vo cancer cell growth. Cyclooxygenase 2(COX-2) is an enzyme that is involved in the conversion of arachidonic acid into prostaglandin E2(PGE2) in humans. PGE2 can promote COX-2 protein expression and tumor cell proliferation and was used as a control.RESULTS Our results showed that 20 μmol/L TQ significantly reduced human LoV o colon cancer cell proliferation. TQ treatment reduced the levels of p-PI3 K, p-Akt, p-GSK3β, and β-catenin and thereby inhibited the downstream COX-2 expression. Results also showed that the reduction in COX-2 expression resulted in a reduction in PGE2 levels and the suppression of EP2 and EP4 activation. Further analysis showed that TG treatment inhibited the nuclear translocation of β-catenin in LoV o cancer cells. The levels of the cofactors LEF-1 and TCF-4 were also decreased in the nucleus following TQ treatment in a dose-dependent manner. Treatment with low dose TQ inhibited the COX-2 expression at the transcriptional level and the regulation of COX-2 expression efficiently reduced LoV o cell migration. The results were further verified in vivo by confirming the effects of TQ and/or PGE2 using tumor xenografts in nude mice.CONCLUSION TQ inhibits LoV o cancer cell growth and migration, and this result highlights the therapeutic advantage of using TQ in combination therapy against colorectal cancer.

Estradiol agonists inhibit human Lo Vo colorectal-cancer cell proliferation and migration through p53

AIM: To investigate the effects of 17β-estradiol via estrogen receptors(ER) or direct administration of ER agonists on human colorectal cancer. METHODS: Lo Vo cells were established from the Bioresource Collection and Research Center and cultured in phenol red-free DMEM(Sigma, United States). To investigate the effects of E2 and/or ER selective agonists on cellular proliferation, Lo Vo colorectal cells were treated with E2 or ER-selective agonists for 24 h and 48 h and subjected to the MTT(Sigma) assay to find the concentration. And investigate the effects of E2 and/or ER selective agonists on cell used western immunoblotting to find out the diversification of signaling pathways. In order to observe motility and migration the wound healing assay and a transwell chamber(Neuro Probe) plate were tased. For a quantitative measure, we counted the number of migrating cells to the wound area post-wounding for 24 h. We further examined the cellular migration-regulating factors urokinase-type plasminogen activator(u-PA), tissue-type plasminogen activator(t-PA) and matrix metalloproteinase(MMP)-9 in human Lo Vo cells so gelatin zymography that we used and gelatinolytic activity was visualized by Coomassie blue staining. And these results are presented as means ± SE, and statistical comparisons were made using Student's t-test.RESULTS: The structure was first compared with E2 and ER agonists. We then treated the Lo Vo cells with E2 and ER agonists(10-8 mol/L) for 24 h and 48 h and subsequently measured the cell viability using MTT assay. Our results showed that treatment with 17β-estradiol and/or ER agonists in human Lo Vo colorectal cancer cells activated p53 and then up-regulated p21 and p27 protein levels, subsequently inhibiting the downstream target gene, cyclin D1, which regulates cell proliferation. Taken together, our findings demonstrate the anti-tumorigenesis effects of 17β-estradiol and/or ER agonists and suggest that these compounds may prove to be a potential alternative therapy in the treatment of human colorectal cancer. These results demonstrate that 17β-estradiol and/or ER agonists downregulate migration-related proteins through the p53 signaling pathway in human Lo Vo colorectal cancer cells. These findings suggest that p53 plays a critical role in the 17β-estradiol and/or ER agonist-mediated protective activity against colorectal cancer progression. In addition, 17β-estradiol and/or ER agonists dramatically inhibited cell migration and reduced the expression of u-PA, t-PA and MMP-9 as well as MMP-2/9 activity in Lo Vo cells, which regulate cell metastasis. Moreover, we observed that pretreatment with a p53 inhibitor significantly blocked the anti-migration effects of E2 and/or ER agonists on Lo Vo cells. That E2 and/or ER agonists may impair Lo Vo cell migration by modulating migration-related factors via the p53 tumor suppressor gene.CONCLUSION: Direct ER treatment may prove to be an attractive alternative therapy in the treatment of human colorectal tumors in the future.

The utilization of traditional Chinese medicine and associated factors in Taiwan in 2002

Background Previous studies have noted that there is a high utilization rate of traditional Chinese medicine （TCM） services in Taiwan, China and in western countries, but few studies investigated factors associated with the utilization of TCM in Taiwan. This study analyzes the utilization rate and the factors associated with the utilization of TCM in Taiwan. Methods Data for this study were from the 2002 HPKAP Survey that conducted the face-to-face questionnaire interviews of people aged 15 years and over from October 2002 to March 2003 in Taiwan. This study analyzed the utilization of TCM outpatient services, including admission to the hospital and clinic visits. Results A total of 26 755 participants completed the survey in the six-month period. The data revealed that 10.4% of participants had utilized TCM services in the past one month while 4.2% of participants utilized TCM only （without using Western medicine outpatient services （WM） or Folk therapy （FT））. The average visits of TCM services per patient was higher among people who had utilized TCM and FT services （2.68 visits） than among those who had utilized WM and FT services （2.15 visits） or TCM services alone （2.15 visits） during the previous one month. Younger people （odds ratio OR = 1.78, 95%CI= 1.47-2.16）, women （compared with men）, and people with higher education levels （OR = 1.58, 95%CI= 1.25-1.98） were more likely to visit TCM than compared groups. People with self-reported poor health status （OR = 2.07, 95%CI= 1.76-2.44） and people who exercise regularly （OR = 1.17, 95%CI= 1.07-1.27） had higher ORs to visit TCM service than comparison group. Conclusions There is a high utilization of TCM in Taiwan. Further studies are needed to investigate the related factors and determinants between the utilization of TCM and the utilization of FT in Taiwan.

The Mechanistic Studies of Acupuncture and Moxibustion in Taiwan

Traditional Chinese acupuncture has a history of over 2500 years. It is effective in the treatment of many conditions with few side effects. The best known mechanism is via endogenous opiates and their receptors. In addition to opioids, researchers have focused on the role of central monoamimergic systems. Acupuncture therapy is used not only to relieve pain but also to treat various medical conditions in traditional Chinese medicine （TCM）. Some experiments have revealed a relationship between acupuncture and the autonomic nervous system （ANS）. Besides, electroacupuncture （EA） can modulate the imbalance between innate and acquired immune systems. This review is focusing on the mechanistic studies of acupuncture that my colleagues and I have performed in Taiwan in recent years. We found that EA analgesia was closely related to not only the serotonergic neurons but also the adrenergic neurons in the central nervous system. The electrophysiological recordings suggested the involvement of the cerebral cortex in acupuncture. Local somatothermal stimulation inhibited the motility of sphincter of Oddi and internal anal sphincter through nitrergic neural release of nitric oxide. Mild local heat stress upregulated hepatic gene expression of heat shock protein 70 and protected the liver from subsequent ischemia- reperfusion injury. These studies supplement the knowledge of the mechanism of acupuncture.

Cycling and Tai Chi Chuan exercises exert greater immunomodulatory effect on surface antigen expression of human hepatitis B virus

Background Both athletes with intensive exercise and aged people may have weakened immunity against virus infection. This study aimed to evaluate whether people undergoing aerobic exercises including competitive cyclists with moderate training （CMT） and middle-aged people practicing Tai Chi Chuan （TCC） exercise have higher immunity against hepatitis B virus than age-matched sedentary controls including college students （CSC） and middle-aged people （MSC）. Methods Human peripheral blood mononuclear cells from competitive cyclists and sedentary controls were stimulated by phytohemagglutinin （PHA） to prepare conditioned medium （MNC-CM） for the assessment of inhibitory effects on hepatitis B surface antigen （HBsAg） expression in human hepatoma Hep3B cells. Results The inhibitory effects on the relative HBsAg expression of CMT＇s and TCC＇s MNC-CM were greater than those of the controls. The CMT＇s MNC-CM prepared from 5 μg/ml PHA decreased HBsAg expression to 61.5%, whereas that of CSC remained at 83.8%. Similarly, this expression by treatment of TCC group＇ MNC-CM was 68.4% whereas that of MSC group was 84.3%. The levels of cytokines such as interferon-γ（IFN-γ）, tumor necrosis factor-α （TNF-α）, IFN-α and interleukin-1β （IL-1β） in the MNC-CM from the CMT and TCC groups were greater than those in the controls. Antibody neutralization of CMT＇s MNC-CM and addition of recombinant cytokines into CSC＇s MNC-CM indicated that IFN-γ, TNF-α and IFN-α had synergistic effects against HBsAg expression. Similar blocking effect was noted in TCC versus MSC groups. Conclusion These results suggest that the immunomodulatory response to suppress HBsAg expression in CMT and TCC with moderate aerobic exercise is greater than that in age-matched sedentary controls.

Effect of auricular acupuncture on oxygen consumption of boxing athletes

Boxing is an official sport at the 2008 Beijing Olympic Games and the fast development of world-class high strength training and sport science has made a significant impact on scientific training. Boxing needs high cardio-respiratory function, speed, muscle strength, and anaerobic and intensive physical demands including weight control covering the grading of athlete＇s body weight and balancing physical ability and health. Sport training is important even for talented athletes. Weight cycling is a term used to describe rapid weight loss following self food limitation and/or dehydration. Both gradual （seasonal） reduction cycles are used and rapid （weekly） weight by athletes,