Modulation of COX2 and hTERT expression by Photodynamic Therapy in human colon cancer cells

Photodynamic therapy (PDT) was employed as a cancer therapy with photosensitizer (PS)-loaded cancer cells, eradicated by the reactive oxygen species after light activation. Cyclo-oxygenase 2 (COX2) is an enzyme expressed in 80% of colon adenocarcinoma and is one of the targets for effective cancer treatment. There is also uprising evidence that the human telomerase reverse transcriptase (hTERT), a catalytic component of telomerase, is reported as a promising indicator for monitoring cancer treatment. In this study, NPe6 mediated PDT on COX2 induced apoptosis in HT-29 was investigated. The cell cycle changes was analysed by flow cytometry and the hTERT expression at pre and post PDT was evaluated at transcription level by Taqman real time PCR. NPe6-PDT in HT-29 cells demonstrated anti-proliferating effect in a drug and light dose dependent manner. LD50 was achieved at 16μg/mL and 2J/cm2 at 4 hour-post treatment with a significant down-regulation of COX2 expression at LD30 and LD50 by immunohistochemical staining (IHC) (p<0.05, One-Way ANOVA). Membrane blebbing was detected in over 60% of cells. 35.2% of treated cells arrested in S-phase at LD50 after 24 hours by flow cytometry. A 0.25- and 0.6-fold down-regulation of hTERT mRNA expression was achieved at LD30 and LD50 respectively by TaqMan real-time PCR. To summarize, NPe6 mediated PDT down-regulated COX2 expression and triggered cell apoptosis. The hTERT can serve as an indicative marker for monitoring NPe6-PDT cancer treatment efficacy.published_or_final_versio

Anti-metastatic mechanism of Tian-Xian Liquid (TXL) and its bioactive fractions in human colorectal cancer cells and xenograft models

Poster Session A: abstract no. 29Colorectal carcinoma is the second most prevalent cancer with an up-rising trend in Hong Kong (Hong Kong Cancer Registry). Traditional Chinese medicine acts as a complementary alternative for tumour therapy with minimal side-effects and traumatic injuries. Tian-Xian Liquid (TXL), one of the well-known natural medicinal herbal formulations, has been commercially used as an anticancer dietary supplement for a decade without known adverse effects. This study aimed to investigate the anti-metastatic property of TXL and its bioactive fractions [butanol fraction (BU), ethyl-acetate fraction (EA) and aqueous fraction (WA)] at molecular level on human colorectal cancer in vitro (HT-29 cancer cells) and in vivo (nude mice xenografts). For the cell model, TXL and its bioactive fractions have similar anti-proliferative effects by MTT assay. At 4-hour-incubation, IC50 values were obtained at 1% (V/V) TXL, 1.25% (V/V) BU, 5% (V/V) EA and 0.3125% (V/V) WA. At IC50, TXL and its bioactive fractions significantly reduced the MMP2 and MMP7 expressions at mRNA level by real-time PCR. At protein level, TXL, BU and EA correspondingly down-regulated MMP2 (active form) and MMP7 protein from 24 to 48 hours; TXL and BU also down-regulated VEGF protein expression; however, no such effect was found in WA-treated cells. Further, only TXL, EA and WA effectively inhibited the cell migration at 48 hours incubation by woundhealing assay. For the xenografts models, MMP2 and MMP7 mRNA expressions were reduced by TXL-, BU- and EA-treated xenografts; however no effects on MMP2 protein expression in all drug-treated xenografts. The VEGF protein expression was significantly down-regulated in TXL- and WA-treated xenografts. Further, TXL, BU and WA effectively inhibited the tumor growth without altering the body weight of the xenografts. In summary, the Chinese medicinal formulation, TXL, demonstrated the most effective anti-metastatic ability on human colorectal cancer in vitro and in vivo.published_or_final_versio

Tian Xian Liquid (TXL) induces apoptosis in HT-29 colon cancer cell in vitro and inhibits tumor growth in vivo

2010-2011 > Academic research: refereed > Publication in refereed journalVersion of RecordPublishe

Photodynamic action of LED-activated nanoscale photosensitizer in nasopharyngeal carcinoma cells

Background: Photodynamic therapy has been confirmed to an efficient therapeutic modality of malignant tumors. The aim of the present study was to explore the photodynamic action of LED-activated nanoscale photosensitizer- loading hypocrellin in nasopharyngeal carcinoma cells. Material/Methods: Nasopharyngeal carcinoma cell line CNE2 cells were subjected to photodynamic therapy with hypocrellinloaded nanophotosensitizer. The uptake of the nanophotosensitizer in the CNE-2 cells was measured using a spectrophotometer and photodynamic toxicity was investigated 18 h after LED radiation treatment. Apoptosis was determined using flow cytometry with propidum iodine staining, and nuclear staining with Hoechst 33258. Active caspase-3 in the CNE2 cells was evaluated using flow cytometry with phycoerythrin (PE)-conjugated anti-active caspase-3 antibodies. Results: The cellular uptake of the nanophotosensitizer in the CNE-2 cells reached optimal at 6 h. LED-activated nanophotosensitizer resulted in doseand light-dependent phototoxicity. Apoptotic rate 18 h after PDT increased to 34.32 ± 1.94% under the light energy of 1 J/cm2. Hoechest 33258 staining reinforced the findings above. Condensation of chromatin and nuclear fragmentations was found in many PDT-treated cells. The activated caspase-3 in the CNE2 cells significantly increased up to 43.90% when the CNE2 cells were exposed to the nanophotosensitizer for 6 h and then 1 J/cm2 irradiation. Conclusion: LED-activated nanophotosensitizer significantly killed the CNE2 cells and enhanced apoptosis and activated caspase-3 in the CNE2 cells. The hypocrellin-loaded nanophotosensitizer might be efficient photosensitizer and LED-activated nanophotosensitizer can be developed for treating nasopharyngeal carcinoma.link_to_subscribed_fulltex

Differential effects of anti-metastatic mechanism of Tian-Xian liquid (TXL) and its bioactive fractions on human colorectal cancer models

Aim of study: This study aimed to elucidate and compare the anti-metastatic mechanism of Tian-Xian liquid (TXL) and its bioactive components namely butanol (BU), ethyl-acetate (EA) and aqueous (WA) fractions on human colorectal cancer in vitro (HT-29 cancer cells) and in vivo (nude mouse xenografts). Materials and methods: The anti-proliferative effects of TXL and its bioactive components in HT-29 cells were determined by MTT assay. Their modulations on the potential angiogenic and metastatic marker expressions on HT-29 cells and xenografts were investigated by real-time PCR and Western blot at transcriptional and translational levels, respectively. For the in vitro study, migration abilities of HT-29 cells were determined using wound healing assay. For the in vivo study, daily measurements of the tumor size and volume of the xenografts were also performed. Results: TXL, BU, EA and WA effectively inhibited the proliferation of HT-29 cells in a dose- and time-dependent manner. The IC 50 value of TXL on HT-29 cells was obtained after incubation with 1% (v/v) TXL for 4 h; whereas IC 50 values were obtained for the following bioactive components: BU at 1.25% (v/v); EA at 5% (v/v); and WA at 0.3125% (v/v). It was found that 1% (v/v) TXL significantly down-regulated MMP2 and MMP7 expression at both transcriptional and translational levels and it reduced MMP9 and VEGF protein expression in vitro. TXL decreased the metastatic ability of HT-29 cells as demonstrated by wound healing assay. TXL and its bioactive fractions caused no significant changes in the body weight indicating lack of toxicity to the xenografts. Conclusions: In summary, TXL multi-targeted to down-regulate the metastatic markers in both in vitro and in vivo models. However, the effects of its bioactive fractions were not obvious. This study profoundly elucidated the anti-proliferative mechanism of TXL, which is vital for the development of future anti-cancer regime in Chinese medicinal formulations. © 2011 Elsevier Ireland Ltd All rights reserved.link_to_subscribed_fulltex

Pharmacological effects of Erxian Decoction, a Chinese medicinal formula, in relieving menopausal syndrome

BACKGROUND: Ovarian aging is an inevitable event since the birth of women. Ovarian aging, together with the senescence of the reproductive axis, will eventually lead women to a transit into menopause, which is defined "after 12 months of amenorrhea following the final menstrual period", according to the STRAW staging system (1). The decrease in ovarian estrogen production is though to related to various menopausal symptoms including hot flash (2), osteoporosis (3), increased risk of cardiovascular diseases (4). Although menopausal syndrome can be relieved by conventional hormone replacement therapy, concerns about the cancer incidence (5) herald a safer yet effective alternative therapy. Erxian Decoction (EXD) has been used for relieving menopausal symptoms for more than fifty years. It is composed of six herbs namely Curculigo orchioides (Hypoxidaceae, rhizome), Epimedium brevicornum (Berberidaceae, whole herb), Morinda officinalis How (Rubiaceae, root), Angelica sinensis (Umbelliferae, root), Phellodendron chinense (Rutaceae bark) and Anemarrhena asphodeloides (Anthericaceae, rhizome). To elucidate the science behind its medical efficacy, intensive effort has been spent by our research group. In this abstract, various pharmacological effects of EXD displayed in vivo or in vitro are discussed. Stimulatory effect of EXD on ovarian estrogen biosynthesis It is found that EXD can increase circulating estrogen level through activation of ovarian aromatase, and thus stimulating ovarian estrogen biosynthesis. Currently, the molecular mechanism of EXD is also being studied on rat ovarian granulosa cell. It is also demonstrated that EXD could also stimulate secretion of estradiol from rat ovarian granulosa cell culture. In twelve-month old female SD-rat fed with EXD, the mRNA level of ovarian aromatase (CYP19) increased significantly, while the serum estradiol level were also significantly elevated, suggesting the stimulatory effect of EXD on, at least, ovarian estrogen biosynthesis (6). Effect of EXD on serum lipid profile In another experiment, the effect of EXD on serum lipid profile has been evaluated. It was demonstrated that EXD could significantly decrease the serum total cholesterol level and serum LDL-cholesterol level. Immunoblotting study revealed that the beneficial effect of EXD on serum lipid profile is at least through up-regulation of hepatic LDL-receptor, a protein responsible for clearance of LDL-cholesterol, and down-regulation of HMG-CoA reductase, a key enzyme for cholesterol synthesis (7) Anti-osteoporotic effect of EXD It is known that the incidence of osteoporosis increased after menopause where circulating estrogen level declines. In our recent study, the anti-osteoporotic effect of EXD has been studied in vitro. It was found that EXD could significantly inhibit the proliferation of RAW264.7, the precursor of osteoclast. On the other hand, EXD could also significantly stimulate the proliferation of hFOB1.19, the human fetal osteoblast cell line. Our results revealed that EXD could increase bone formation and reduce bone resorption through regulating the respective cell proliferation. Results from TRAP staining of RAW264.7 also revealed that EXD could inhibit osteoclastogenesis through inhibition of RAW264.7 differentiation. EXD could also increase the trabecular material bone mineral density at the L2 mid-vertebral body in 12-month old female SD-rat, as revealed by microcomputed tomography, suggesting its anti-osteoporotic property in vivo (8). Anti-oxidative effect of EXD EXD also possess anti-oxidative properties, as revealed by the mRNA level of hepatic anti-oxidative enzymes SOD, CAT and GPx-1 (6). Since aging is thought to relate with oxidative stress, the stimulatory effect of EXD on anti-oxidative enzymes suggests the general anti-aging properties of EXD and merits further study. CONCLUSION: The research of diverse pharmacological effects of EXD in vivo and in vitro has been discussed. With the stimulatory effect of EXD on ovarian estrogen biosynthesis, beneficial effects of EXD on serum lipid profile, anti-osteoporotic properties together with the anti-oxidative effects, it is promising that EXD could be an effective alternative for relieving menopausal syndromes.The 13th World Congress on Menopause (IMS Roma 2011), Rome, Italy, 8-11 June 2011