Toona Sinensis and Moschus Decoction Induced Cell Cycle Arrest in Human Cervical Carcinoma HeLa Cells Hong Zhen,1 Yifei Zhang,1 Zhijia Fang,2 Zhiwei Huang,2 Chongge You,3 and Ping Shi1
1State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, 130 Meilong Road, Shanghai 200237, China 2College of Chemistry, Chemical Engineering and Biotechnology, Donghua University, 2999 Renmin Road, Shanghai 201620, China 3Lanzhou University Second Hospital, 82 Cuiying Gate, Lanzhou 730030, China
Toona sinensis and Moschus are two herb materials used in traditional Chinese medicine, most commonly for their various biological activities. In this study, we investigated the inhibitory effect of three decoctions from Toona sinensis, Moschus, and Toona sinensis and Moschus in combination on cell growth in several normal and cancer cell lines by cell viability assay. The results showed that the combined decoction exhibited the strongest anticancer effects, compared to two single decoctions. The observations indicated that the combined decoction did not induce cell apoptosis and autophagy in HeLa cells by fluorescence microscopy. Flow cytometry analysis revealed that the combined decoction arrested HeLa cell cycle progression in S-phase. After the decoction incubation, among 41 cell cycle related genes, eight were reduced, while five were increased in mRNA levels by real-time PCR assay. Western blotting showed that there were no apparent changes of protein levels of Cyclin E1, while P27 expression significantly declined and the levels of CDC7 and CDK7 obviously increased. The data suggest that the RB pathway is partially responsible for the decoction-induced S-phase cell cycle arrest in HeLa cells. Therefore, the combined decoction may have therapeutic potential as an anticancer formula for certain cancers. Conclusion This study is the first to report the anticancer activity of decoction of Toona sinensis and Moschus in combination. Furthermore, we have shown that the Toona sinensis and Moschus decoction exhibits its anticancer effects by inducing cell cycle arrest in S-phase via mRNA regulation of cell cycle related proteins in HeLa cells. Our data provide strong evidence that the Toona sinensis and Moschus decoction could be potentially utilized for the treatment of some cancers.
Berberine modulates AP-1 activity to suppress HPV transcription and downstream signaling to induce growth arrest and apoptosis in cervical cancer cells Sutapa Mahata1, Alok C Bharti1, Shirish Shukla1, Abhishek Tyagi1, Syed A Husain2 and Bhudev C Das13*
1 Division of Molecular Oncology, Institute of Cytology and Preventive Oncology (Indian Council of Medical Research), I-7, Sector-39, Noida, Gautam Budh Nagar - 201301 India 2 Department of Biosciences, Faculty of Natural Sciences, Jamia Millia Islamia, New Delhi -110025, India 3 Dr. B.R Ambedkar Centre for Biomedical Research, University of Delhi (North Campus), Delhi-110007, India
Background- Specific types of high risk Human papillomaviruses (HR-HPVs) particularly, HPV types 16 and 18 cause cervical cancer and while the two recently developed vaccines against these HPV types are prophylactic in nature, therapeutic options for treatment and management of already existing HPV infection are not available as yet. Because transcription factor, Activator Protein-1 (AP-1) plays a central role in HPV-mediated cervical carcinogenesis, we explored the possibility of its therapeutic targeting by berberine, a natural alkaloid derived from a medicinal plant species, Berberis which has been shown to possess anti-inflammatory and anti-cancer properties with no known toxicity; however, the effect of berberine against HPV has not been elucidated.
Results- We studied the effect of berberine on HPV16-positive cervical cancer cell line, SiHa and HPV18-positive cervical cancer cell line, HeLa using electrophoretic mobility gel shift assays, western and northern blotting which showed that berberine could selectively inhibit constitutively activated AP-1 in a dose- and time-dependent manner and downregulates HPV oncogenes expression. Inhibition of AP-1 was also accompanied by changes in the composition of their DNA-binding complex. Berberine specifically downregulated expression of oncogenic c-Fos which was also absent in the AP-1 binding complex. Treatment with berberine resulted in repression of E6 and E7 levels and concomitant increase in p53 and Rb expression in both cell types. Berberine also suppressed expression of telomerase protein, hTERT, which translated into growth inhibition of cervical cancer cells. Interestingly, a higher concentration of berberine was found to reduce the cell viability through mitochondria-mediated pathway and induce apoptosis by activating caspase-3.
Conclusion- These results indicate that berberine can effectively target both the host and viral factors responsible for development of cervical cancer through inhibition of AP-1 and blocking viral oncoproteins E6 and E7 expression. Inhibition of AP-1 activity by berberine may be one of the mechanisms responsible for the anti-HPV effect of berberine. We propose that berberine is a potentially promising compound for the treatment of cervical cancer infected with HPV.
Anticancer Activity of Certain Herbs and Spices on the Cervical Epithelial Carcinoma (HeLa) Cell Line Danielle Berrington and Namrita Lall
Department of Plant Sciences, University of Pretoria, Pretoria 0002, South Africa
Abstract Acetone extracts of selected plant species were evaluated for their in vitro cytotoxicity against a noncancerous African green monkey kidney (Vero) cell line and an adenocarcinoma cervical cancer (HeLa) cell line. The plants studied were Origanum vulgare L. (Oregano), Rosmarinus officinalis L. (Upright and ground cove rosemary), Lavandula spica L. (Lavender), Laurus nobilis L. (Bay leaf), Thymus vulgaris L. (Thyme), Lavandula x intermedia L. (Margaret Roberts Lavender), Petroselinum crispum Mill. (Curly leaved parsley), Foeniculum vulgare Mill. (Fennel), and Capsicum annuum L. (Paprika). Antioxidant activity was determined using a quantitative DPPH (1,1-diphenyl-2-picryl hydrazyl) assay. The rosemary species exhibited effective radical scavenging capacity with 50% inhibitory concentration (IC50) of 3.48 +/- 0.218 μg/mL and 10.84 +/- 0.125 ug/mL and vitamin C equivalents of 0.351 g and 1.09 g for McConnell’s Blue and Tuscan Blue, respectively. Cytotoxicity was measured using XTT (Sodium 3'-1-(phenyl amino-carbonyl)-3,4-tetrazolium]-bis-[4-methoxy-6-nitro] benzene sulfonic acid hydrate) colorimetric assay. Only L. nobilis and O. vulgare exhibited pronounced effects on the HeLa cell line. Dose-dependent studies revealed IC50 of 34.46 μg/mL and 0.48 μg/mL on the HeLa cells and on the Vero cells 124.1 μg/mL ± 18.26 and 163.8 μg/mL ± 2.95 for L. nobilis and O. vulgare, respectively. Light (eosin and haematoxylin staining) and confocal microscopy (Hoechst 33342, acridine orange, and propidium iodide staining) were used to evaluate the cytotoxic mechanism of action for L. nobilis and O. vulgare.
Conclusion In this study common herbs and spices were chosen to determine their anticancer activity. Herbs and spices were chosen due to their bioactive components which have the ability to reduce the risk of cancer through their antimicrobial, antioxidant, and antitumourogenic activity . The rosemary species were found to have the highest antioxidant content which could be attributed to the high content of polyphenolic compounds. Therefore the rosemary species possess potential chemopreventive properties. After performing the XTT cytotoxicity assay it was hypothesized that Laurus nobilis, with an SI of 3.61, and Oregano vulgare, with an SI of 1.30, were the best candidates for further investigation. Both these extracts were further investigated to determine their mechanism of cell death by observing them under light microscopy and confocal microscopy. From the microscopy images it was observed that in both cell lines morphological changes did appear after exposure to various concentrations of the acetone extracts. Although these microscopy images were in agreement with one another that apoptosis was the possible mechanism of cell death, as seen by the characteristic features of hypercondensed chromatin (mostly in HeLa cells) and cellular debris (seen only in Vero cells), they are not sufficient enough to definitely confirm that apoptosis is taking place and therefore more sensitive assays such as flow cytometry need to Be used to confirm that apoptosis is taking place. However Laurus nobilis did show promising results as an anticancer agent due to its relatively high toxicity on the HeLa cell line and at this same concentration a low toxicity on the Vero cell line.
Phytochemicals, antioxidant properties and anticancer investigations of the different parts of several gingers species (Boesenbergia rotunda, Boesenbergia pulchella var attenuata and Boesenbergia armeniaca) Ling Jing Jing1, Maryati Mohamed2, Asmah Rahmat3 and Mohd Fadzelly Abu Bakar1*
1Laboratory of Natural Products, Institute for Tropical Biology and Conservation, Universiti Malaysia Sabah, Locked Bag No. 2073, 88999, Kota Kinabalu, Sabah, Malaysia. 2Faculty of Civil Engineering and Environment, Universiti Tun Hussein Onn Malaysia, 86400 Parit Raja, Batu Pahat, Johor, Malaysia. 3Department of Nutrition and Dietetics, Faculty of Medicine and Health Sciences, Universiti Putra Malaysia, 43400, UPM, Serdang, Selangor, Malaysia.
Abstract Extracts (methanol) of the leaves, stem and rhizome of Boesenbergia species were studied for their phytochemical constituents, total phenolics and flavonoid contents, antioxidant as well as anticancer properties. The plants revealed the presence of polyphenols such as quercetin, kaempferol, rutin, naringin, hesperidin, caffeic acid, p-coumaric acid, ferulic acid, sinapic acid, chlorogenic acid, gallic acid, luteolin and diosmin by using High Performance Liquid Chromatographic (HPLC). It was indicated with significant composition of hesperidin and naringin in B. pulchella var attenuata (leaves and stem); quercetin and kaempferol in B. rotunda; luteolin in B. armeniaca. The results of antioxidant assessments conducted were similar to the trend of total phenolic and flavonoid contents: B. pulchella var attenuata> B. rotunda> B. armeniaca. In the cytotoxicity assay, B. rotunda showed the most prominent and promising result as anticancer medicinal plant. It showed positive antiproliferative effect against five cancer cell lines: ovarian (CaOV3), breast (MDA-MB-231 and MCF-7), cervical (HeLa) and colon (HT-29) cancer cell lines with 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay conducted. In addition, the rhizome of B. pulchella var attenuata and B. armeniaca shown positive result in cytotoxicity assay tested against breast cancer (MCF-7). Thus, the Boesenbergia species investigated would be a promising anticancer remedy for breast cancer.
Source : Journal of Medicinal Plants Research Vol. 4(1), pp. 027-032, 4 January, 2010 Link to Full Article
Antiproliferative activity of different extracts from Daphne altaica Pall. on selected cancer cells Murat Kizaibek1,3, Marzia Daniar2, Lin Li2 and Halmurat Upur3*
1Traditional Kazakh Medicine Research Institute of Ili Kazakh Autonomous Prefecture, 835000 Yining, Xinjiang, P. R. China. 2Department of Biochemistry and Molecular Biology, College of Basic Medicine, Xinjiang Medical University, 830011 Urumqi, Xinjiang, P. R. China. 3Faculty of Traditional Uighur Medicine, Xinjiang Medical University, 830011 Urumqi, Xinjiang, P. R. China.
Abstract Daphne altaica Pall. (Thymelaeaceae) is a medicinal plant that has long been used to treat cancer and respiratory ailments in Traditional Kazakh Medicine. In order to systematically evaluate its potential anticancer activity, six extracts of different polarity, namely: aqueous, n-butanol, ethyl acetate, chloroform, petroleum ether and ethanol extracts were obtained from this plant and they were tested for their antiproliferative effects on four human cancer cell lines: esophageal squamous cell carcinoma, gastric carcinoma, hepatoma and cervical carcinoma cells. Results from the proliferation assay showed that all extracts, except for aqueous extract, exhibited a dose-dependent growth inhibitory effect on all cancer cell lines. Of these extracts, two fractions obtained from partitioning of ethanol extract, namely: chloroform extract and ethyl acetate extract, could be considered a potential source of anticancer compounds. Further studies are necessary for identification and chemical characterization of the active principles.
Conclusion The results of this study represent the first evidence that barks of D. altaica possess effective antiproliferative activities on human cancer cells. Future phytochemical investigations on this medicinal plant should be encouraged.
Source : Journal of Medicinal Plants Research Vol. 5(15), pp. 3448-3452, 4 August, 2011 Link to Full Article
Inhibition of Human Cervical Cancer Cell Growth by Ethanolic Extract of Boerhaavia diffusa Linn. (Punarnava) Root Rakhi Srivastava,1 Daman Saluja,1 Bilikere S. Dwarakanath,2 and Madhu Chopra1
1Dr. B. R. Ambedkar Center for Biomedical Research, University of Delhi, Delhi 110007, India 2Institute of Nuclear Medicine and Allied Sciences, Delhi 110054, India
Abstract In Indian traditional medicine, Boerhaavia diffusa (punarnava) roots have been widely used for the treatment of dyspepsia, jaundice, enlargement of spleen, abdominal pain and as an anti-stress agent. Pharmacological evaluation of the crude ethanolic extract of B. diffusa roots has been shown to possess antiproliferative and immunomodulatory properties. The extract of B. diffusawas studied for anti-proliferative effects on the growth of HeLa cells and for its effect on cell cycle. Bio-assays of extracts from B. diffusa root showed that a methanol : chloroform fraction (BDF 5) had an antiproliferative effect on HeLa cells. After 48 h of exposure, this fraction at a concentration of 200 μg mL−1 significantly reduced cell proliferation with visible morphological changes in HeLa cells. Cell cycle analysis suggests that antiproliferative effect of BDF 5 could be due to inhibition of DNA synthesis in S-phase of cell cycle in HeLa cells, whereas no significant change in cell cycle was detected in control cells. The fraction BDF 5 caused cell death via apoptosis as evident from DNA fragmentation and caspase-9 activation. Thus the extract has potential to be evaluated in detail to assess the molecular mechanism-mediated anticancer activities of this plant.
Source Evidence-Based Complementary and Alternative Medicine Volume 2011 (2011) Link to source
A novel polysaccharide, isolated from Angelica sinensis (Oliv.) Diels induces the apoptosis of cervical cancer HeLa cells through an intrinsic apoptotic pathway.
Cao W, Li XQ, Wang X, Fan HT, Zhang XN, Hou Y, Liu SB, Mei QB. Department of Pharmacology, School of Pharmacy, the Fourth Military Medical University, 169 West Changle Road, Xi'an, Shaanxi 710032, China.
A novel polysaccharide isolated from Angelica sinensis, named APS-1d showed cytotoxic activity towards several cancer cell lines in vitro. However, the precise antitumor mechanisms of this compound are unknown. In this study, we investigated the pro-apoptotic effects of APS-1d in human cervical cancer HeLa cells both in vitro and in vivo, and further elucidated the mechanisms of this action. Inhibition of HeLa cell proliferation was determined by MTT assay and the therapeutic efficacy of APS-1d was evaluated by human cancer xenografts in nude mice. Cell apoptosis was examined with flow cytometry and TUNEL assay. The mechanism of action of APS-1d was investigated by Western blot analysis. APS-1d decreased HeLa cell proliferation in a concentration- and time-dependent manner in vitro. In addition, APS-1d significantly inhibited tumor growth in athymic nude mice. Characteristic manifestations of apoptosis including apoptotic morphological features and the sub- [G.sub.0]/[G.sub.1] peaks were observed when the cells were treated with APS-1d. Further analysis showed that APS-1d-induced apoptosis was associated with the regulation of Bcl-2 family protein expression, a decrease in the mitochondrial membrane potential, and an increase in the cytosolic cytochrome c levels. Sequentially, APS-1d increased the activities of caspase-9, -3, and poly (ADP-ribose) polymerase in a concentration-dependent manner, however, no obvious activation of Bid and caspase-8 was observed. Pretreatment with Z-LEHD-FMK, a specific inhibitor of caspase-9, significantly attenuated APS-1d-induced cell apoptosis, and activation of caspase-3. Taken together, our studies indicate that APS-1d is capable of inhibiting HeLa cell proliferation and inducing apoptosis in these cells which primarily involves the activation of the intrinsic mitochondrial pathway.
Cytotoxic activity of proteins isolated from extracts of Corydalis cava tubers in human cervical carcinoma HeLa cells Robert Nawrot, Maria Wolun-Cholewa,Wojciech Bialas, Danuta Wyrzykowska, Stanislaw Balcerkiewicz and Anna Gozdzicka-Jozefiak
Background Corydalis cava Schweigg. & Koerte, the plant of numerous pharmacological activities, together with the studied earlier by our group Chelidonium majus L. (Greater Celandine), belong to the family Papaveraceae. The plant grows in Central and South Europe and produces the sizeable subterraneous tubers, empty inside, which are extremely resistant to various pathogen attacks. The Corydalis sp. tubers are a rich source of many biologically active substances, with the extensive use in European and Asian folk medicine. They have analgetic, sedating, narcotic, anti-inflammatory, anti-allergic and anti-tumour activities. On the other hand, there is no information about possible biological activities of proteins contained in Corydalis cava tubers.
Methods Nucleolytic proteins were isolated from the tubers of C. cava by separation on a heparin column and tested for DNase activity. Protein fractions showing nucleolytic activity were tested for cytotoxic activity in human cervical carcinoma HeLa cells. Cultures of HeLa cells were conducted in the presence of three protein concentrations: 42, 83 and 167 ng/ml during 48 h. Viability of cell cultures was appraised using XTT colorimetric test. Protein fractions were separated and protein bands were excised and sent for identification by mass spectrometry (LC-ESI-MS/MS).
Results The studied protein fractions showed an inhibiting effect on mitochondrial activity of HeLa cells, depending on the administered dose of proteins. The most pronounced effect was obtained with the highest concentration of the protein (167 ng/ml) - 43.45 +/- 3% mitochondrial activity of HeLa cells were inhibited. Mass spectrometry results for the proteins of applied fractions showed that they contained plant defense- and pathogenesis-related (PR) proteins.
Conclusions The cytotoxic effect of studied proteins toward HeLa cell line cells has been evident and dependent on increasing dose of the protein. The present study, most probably, represents the first investigations on the effect of purified PR proteins from tuber extracts of a pharmacologically active plant on cell lines.
Source : BMC Complementary and Alternative Medicine 2010, 10:78doi:10.1186/1472-6882-10-78 LINK TO FULL ARTICLE
Cinnamon extract induces tumor cell death through inhibition of NFκB and AP1
Background Cinnamomum cassia bark is the outer skin of an evergreen tall tree belonging to the family Lauraceae containing several active components such as essential oils (cinnamic aldehyde and cinnamyl aldehyde), tannin, mucus and carbohydrate. They have various biological functions including anti-oxidant, anti-microbial, anti-inflammation, anti-diabetic and anti-tumor activity. Previously, we have reported that anti-cancer effect of cinnamon extracts is associated with modulation of angiogenesis and effector function of CD8+ T cells. In this study, we further identified that anti-tumor effect of cinnamon extracts is also link with enhanced pro-apoptotic activity by inhibiting the activities NFκB and AP1 in mouse melanoma model.
Methods Water soluble cinnamon extract was obtained and quality of cinnamon extract was evaluated by HPLC (High Performance Liquid Chromatography) analysis. In this study, we tested anti-tumor activity and elucidated action mechanism of cinnamon extract using various types of tumor cell lines including lymphoma, melanoma, cervix cancer and colorectal cancer in vitro and in vivo mouse melanoma model.
Results Cinnamon extract strongly inhibited tumor cell proliferation in vitro and induced active cell death of tumor cells by up-regulating pro-apoptotic molecules while inhibiting NFκB and AP1 activity and their target genes such as Bcl-2, BcL-xL and survivin. Oral administration of cinnamon extract in melanoma transplantation model significantly inhibited tumor growth with the same mechanism of action observed in vitro.
Conclusion Our study suggests that anti-tumor effect of cinnamon extracts is directly linked with enhanced pro-apoptotic activity and inhibition of NFκB and AP1 activities and their target genes in vitro and in vivo mouse melanoma model. Hence, further elucidation of active components of cinnamon extract could lead to development of potent anti-tumor agent or complementary and alternative medicine for the treatment of diverse cancers.
SOURCE : BMC Cancer 2010, 10:392doi:10.1186/1471-2407-10-392 LINK TO FULL ARTICLE Aqueous Cinnamon Extract (ACE-c) from the bark of Cinnamomum cassia causes apoptosis in human cervical cancer cell line (SiHa) through loss of mitochondrial membrane potential
Background Chemoprevention, which includes the use of synthetic or natural agents (alone or in combination) to block the development of cancer in human beings, is an extremely promising strategy for cancer prevention. Cinnamon is one of the most widely used herbal medicines with diverse biological activities including anti-tumor activity. In the present study, we have reported the anti-neoplastic activity of cinnamon in cervical cancer cell line, SiHa.
Methods The aqueous cinnamon extract (ACE-c) was analyzed for its cinnamaldehyde content by HPTLC analysis. The polyphenol content of ACE-c was measured by Folin-Ciocalteau method. Cytotoxicity analysis was performed by MTT assay. We studied the effect of cinnamon on growth kinetics by performing growth curve, colony formation and soft agar assays. The cells treated with ACE-c were analyzed for wound healing assay as well as for matrix metalloproteinase-2 (MMP-2) expression at mRNA and protein level by RT-PCR and zymography, respectively. Her-2 protein expression was analyzed in the control and ACE-c treated samples by immunoblotting as well as confocal microscopy. Apoptosis studies and calcium signaling assays were analyzed by FACS. Loss of mitochondrial membrane potential (Δψm) in cinnamon treated cells was studied by JC-1 staining and analyzed by confocal microscopy as well as FACS.
Results Cinnamon alters the growth kinetics of SiHa cells in a dose-dependent manner. Cells treated with ACE-c exhibited reduced number of colonies compared to the control cells. The treated cells exhibited reduced migration potential that could be explained due to downregulation of MMP-2 expression. Interestingly, the expression of Her-2 oncoprotein was significantly reduced in the presence of ACE-c. Cinnamon extract induced apoptosis in the cervical cancer cells through increase in intracellular calcium signaling as well as loss of mitochondrial membrane potential.
Conclusion Cinnamon could be used as a potent chemopreventive drug in cervical cancer.
Cervical cancer is the second most common carcinoma in the world among women and is highly chemoresistant and radio resistant, often resulting in local treatment failure. For locally advanced disease, radiation is combined with low-dose chemotherapy; however, this modality often leads to severe toxicity. Prevention of cancer through dietary intervention recently has received an increasing interest, and dietary agents have become not only important potential chemopreventive, but also therapeutic agents when combined with chemotherapy or radiotherapy. In this study, we observed that gemcitabine was highly cytotoxic to both cancer and normal cells while clove extract (0.7-8 mg mL-1) was found to be comparatively more cytotoxic towards cancer cells. Notably, combination of low dose gemcitabine and ethanolic clove extract (2 and 3 mg mL-1) had more pronounced cytotoxic effect on cancer cells than single modalities. It is noteworthy that use of clove extract increased the efficacy of gemcitabine and importantly, it was found to be minimally toxic to normal cells. Together, these results suggest a novel mechanism may be involved in the synergistic effect of this combination.