Evaluation of Therapeutic Potential of Selected Plant-Derived Homeopathic Medicines for their Action against Cervical Cancer.

BACKGROUND: Plant-derived homeopathic medicines (HMs) are cheap and commercially available but are mechanistically less explored entities than conventional medicines. PURPOSE: The aim of our study was to evaluate the impact of selected plant-derived HMs derived from Berberis aquifolium (BA), Berberis vulgaris (BV), Mentha piperita (MP), Curcuma longa (CL), Cinchona officinalis (CO), Thuja occidentalis (TO) and Hydrastis canadensis (HC) on cervical cancer (CaCx) cells in vitro. METHODS: We screened the mother tincture (MT) and 30C potencies of the above-mentioned HMs for anti-proliferative and cytotoxic activity on human papillomavirus (HPV)-negative (C33a) and HPV-positive CaCx cells (SiHa and HeLa) by MTT assay. Total phenolic content (TPC) and the free-radical scavenging activity of each HM was also determined using standard assays. Phytochemicals reportedly available in these HMs were examined for their potential inhibitory action on HPV16 E6 by in silico molecular docking. RESULTS: All tested MTs induced a differential dose-dependent cytotoxic response that varied with cell line. For C33a cells, the order of response was TO > CL > BA > BV > HC > MP > CO, whereas for SiHa and HeLa cells the order was HC > MP > TO > CO > BA > BV > CL and CL > BA > CO, respectively. 30C potencies of all HMs showed an inconsistent response. Further, anti-CaCx responses displayed by MTs did not follow the order of an HM's phenolic content or free radical scavenging activity. Analysis revealed anti-oxidant content of BA, BV and HC had the lowest contribution to their anti-CaCx activity. Using in silico modeling of molecular docking between the HPV16 E6 protein crystallographic structures (6SJA and 4XR8) and main phytochemical components of BV, BA, HC, CL and TO, their potential to inhibit the HPV16 E6 protein carcinogenic interactions was identified. CONCLUSION: The study has shown a comparative evaluation of the potential of several plant-derived MTs and HMs to affect CaCx cell line survival in vitro (through cytotoxicity and free radical scavenging) and their theoretical molecular targets in silico for the first time. Data demonstrated that MTs of BA and BV are likely to be the most potent HMs that strongly inhibited CaCx growth and have a strong anti-HPV phytochemical constitution.

In Vitro Assessment of Homeopathic Potencies of Hydrastis canadensis on Hormone-Dependent and Independent Breast Cancer.

BACKGROUND: Breast cancer is the second leading cause of cancer-related deaths in women. Conventional treatment such as chemotherapy, hormonal therapy and radiotherapy has decreased the mortality rate among cancer patients but has also revealed long-term side effects. Drug resistance and toxicity to normal cells compound the problems associated with the use of modern medicines. Hence, complementary or alternative treatment options are being explored. The current study, using different homeopathic potencies of Hydrastis canadensis, was conducted to distinguish between any effects they might have on hormone-dependent and independent breast cancer. MATERIALS AND METHODS: The cytotoxic effect of homeopathic medicine Hydrastis on hormone-dependent (MCF 7) and hormone-independent (MDA-MB-468) breast cancer cells was assessed using viability and colony-forming assays after 48 or 72 hours of treatment. Flow cytometry-based Annexin V-PI (propidium iodide), caspase 3 and cell cycle analysis was performed following treatment of cells with mother tincture or various potencies of Hydrastis (1C, 2C, 30C, 200C). RESULTS: Different potencies of Hydrastis displayed selective cytotoxic effects against MCF 7 cells, but only marginal effects against MDA-MB-468. The maximum cytotoxicity was established in the case of 1C following 72 hours of treatment. Treatment of breast cancer cells revealed an increase in the G0/G1 cell population, along with an increase in the caspase 3 levels and induction of apoptosis. CONCLUSION: Hydrastis may have a selective cytotoxic effect against hormone-dependent breast cancer MCF 7 cells, leading to cell cycle arrest in the G0/G1 phase, which could be the plausible reason for the induction of apoptosis. The results need to be validated in vivo.

Evidence that Extreme Dilutions of Paclitaxel and Docetaxel Alter Gene Expression of In Vitro Breast Cancer Cells.

BACKGROUND: Gene expression analysis of cells treated with extreme dilutions or micro amounts of drugs has been used to provide useful suggestions about biological responses. However, most of the previous studies were performed on medicines being prepared from a variety of herbal and metal sources. This study investigated the effects of ultramolecular dilution of the taxane anti-cancer drugs, which are not commonly used in homeopathic medicines, on mRNA expression profiles of five key genes (p53, p21, COX-2, TUBB2A and TUBB3) in the breast cancer cell line MCF-7. METHOD: MCF-7 cells were exposed to paclitaxel (Taxol) or docetaxel (Taxotere) preparations (6X, 5C and 15C dilutions prepared from pharmacological concentration of 25 nmol/L) for 72 hours. The cell culture groups were evaluated with the trypan blue dye exclusion method for the proliferation/cytotoxicity rates, immuno-staining ß-tubulin for microtubule organization, and reverse transcription polymerase chain reaction for gene expression levels.Fold-change in gene expression was determined by the ΔΔCt method. RESULTS: The administration of diluted preparations had little or no cytotoxic effect on MCF-7 cells, but altered the expression of genes analyzed with a complex effect. According to the ΔΔCt method with a five-fold expression difference (p < 0.05) as a cut-off level, ultra-high dilutions of paclitaxel and docetaxel showed differential effects on the studied genes with a concentration-independent activity. Furthermore, the dilutions disrupted the microtubule structure of MCF-7 cells, suggesting that they retain their biological activity. CONCLUSION: Despite some limitations, our findings demonstrate that gene expression alterations also occur with ultra-high dilutions of taxane drugs.

Effects of ultra-high dilutions of sodium butyrate on viability and gene expression in HEK 293 cells.

BACKGROUND: Several recent studies reported the capability of high diluted homeopathic medicines to modulate gene expression in cell cultures. In line with these studies, we examined whether ultra-high dilutions (30C and 200C) of sodium butyrate (SB) can affect the expression levels of genes involved in acquisition of a senescence-associated secretory phenotype (SASP) in human embryonic kidney (HEK) 293 cells. METHODS: Cell viability was evaluated using a 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) assay. The expression levels of TNF-α, interleukin (IL)-2, IL-4, IL-6 and IL-10 genes were determined by real-time PCR assay. RESULTS: Exposure to both 30C and 200C during 48 h led to a significant decrease of the level of expression of TNF-α gene, while expression of IL-2 gene was increased when exposed to 30C, and expression of IL-10 gene was decreased when exposed to 200C. No changes in expression levels of all genes studied were observed in cells treated with both 30C and 200C remedies of SB during the 24 h. CONCLUSION: Observed changes in gene expression levels after exposure to 30C and 200C remedies of SB during 48 h suggest that extremely low concentrations of this agent can modulate the transcriptome of HEK 293 cells. These results are in line with findings from other studies confirming the ability of homeopathic remedies to modulate gene expression in cell cultures.

Therapeutic potential of HIV nosode 30c as evaluated in A549 lung cancer cells.

OBJECTIVES: To examine if HIV nosode in 30c dilution (HIV 30c) has therapeutic potential against lung cancer cells (A549) as compared to WRL-68 normal cells and to elucidate its possible molecular mechanism of action on DNA replication and apoptosis. METHODS: Effects of HIV 30c were thoroughly tested for its possible anticancer potential on A549 cells (lung cancer); WRL-68 normal liver cells served as control. Three doses, one at LD50 and two below LD-50, were used. Proliferation, migration and senescence assays were made and generation of reactive oxygen species (ROS) studied by routine techniques. The ability of HIV 30c to induce apoptosis in A549 cells and its possible signalling pathway were determined using immunoblots of relevant signal proteins and confocal microscopy, including studies on telomerase reverse transcriptase (TERT) and topoisomerase II (Top II) activities, intimately associated with cell division and DNA replication. RESULTS: HIV 30c prevented cancer cell proliferation and migration, induced pre-mature senescence, enhanced pro-apoptotic signal proteins like p53, bax, cytochrome c, caspase-3 and inhibited anti-apoptotic signal proteins Bcl2, TERT and Top II, changed mitochondrial membrane potential and caused externalization of phosphatidyl serine. Thus, it induced apoptosis as also evidenced from increase in cells with distorted membrane morphology, nuclear condensation, DNA fragmentation, and ROS, typical of apoptosis in progress. CONCLUSION: HIV 30c nosode has therapeutic potential for inducing cytotoxic effects on A549 cells as manifested by changes in nuclear condensation, DNA fragmentation, ROS generation and MMP, and for its inhibitory action on cell proliferation, cell migration, expression of telomerase reverse transcriptase and Top II genes, and increasing expression of pro-apoptotic genes.

Pro-apoptotic and anti-angiogenic properties of the α /ß-thujone fraction from Thuja occidentalis on glioblastoma cells.

The most aggressive type of brain tumor is glioblastoma multiforme, which to date remains incurable. Thuja occidentalis is used in homeopathy for the treatment of cancer, however, its mechanism of action remains unknown. We set out to study the effects of thujone fractions of Thuja on glioblastoma using in vitro and in vivo models. We found that the α/ ß-thujone fraction decrease the cell viability and exhibit a potent anti-proliferative, pro-apoptotic and anti-angiogenic effects in vitro. In vivo assays showed that α /ß-thujone promotes the regression of neoplasia and inhibits the angiogenic markers VEGF, Ang-4 and CD31 into the tumor.

Homeopathic remedies with antineoplastic properties have immunomodulatory effects in experimental animals.

BACKGROUND: Our previous work suggests that Thuja occidentalis, Carcinosinum and Ruta graveolens have antineoplastic properties. The mechanism of this action has not previously been studied. We studied the hypothesis that the mechanism of action is through the immune modulation. METHODS: We evaluated the effects of Thuja occidentalis, Carcinosinum and Ruta graveolens 1M, 200c and 30c on the immune system of Balb/c mice. The homeopathic preparations were administered orally for ten consecutive days. Haematological parameters (Total White Blood Cell (WBC) Count, Differential Count and Haemoglobin content), haematopoietic parameters (bone marrow cellularity and α-esterase positive cells) and immune parameters for antibody response and lymphoid cell proliferation were assessed using standard methods. Results were analysed by statistical comparison with the control. RESULTS: We observed significant enhancement of haematological parameters including total WBC count, haematopoietic parameters such as bone marrow cellularity and the number of α-esterase positive cells, other parameters of immune response such as circulating antibody titre and the number of plaque forming cells (PFC), particularly with higher dilutions of Thuja and Ruta. Enhanced proliferation of B and T lymphoid cells was also observed. No toxic effects were observed. CONCLUSIONS: The results suggest immunomodulatory activity of homeopathic preparations in high dilution. This may be a mechanism through which homeopathic preparations act.

[Research on therapeutic effects of bufonis venenum on L7212 leukemia and its mechanism].

OBJECTIVE: To study the therapeutic effects of Bufonis Venenum on L7212 leukemia and the potential mechanism. METHODS: L7212 leukemia model mice were randomly divided into four groups: model group, low and high dose Bufonis Venenum groups and chemotherapy group. Normal mice were treated as control group. Mice were injected intraperitoneally for 10 days continually. The body weight, survival time, peripheral blood leukocyte, hepatic and splenic indexes, bone marrow leukocyte and T lymphocyte were observed and determined. RESULTS: Body weight of L7212 leukemia model group mice were decreased significantly. Compared with other groups, high dose Bufonis Venenum group's weight loss was the least. Bufonis Venenum groups survived longer than L7212 model group. Compared with model group, high dose Bufonis Venenum group's liver index was higher (P < 0.05). After inoculation for 1 day, leukocyte count as well as percentage of leukemic cells within five groups had no significant difference (P > 0.05). Compared with the model group, after inoculation for 10 days, leukocyte count in Bufonis Venenum groups and the chemotherapy group were significantly reduced (P < 0.05). Percentage of leukemia cells in blood and bone marrow in high dose Bufonis Venenum group was significantly decreased (P < 0.05). Compared with model group, CD3+ and CD4+ in Bufonis Venenum groups and the chemotherapy group were increased, CD8+ was decreased, but had no significant difference (P > 0.05). CONCLUSION: Bufonis Venenum has therapeutic effects on the L7212 leukemia by inducing apoptosis and improving immune system.

Anticancer potential of homeopathic medicines: an updated review.

Globally, cancer is the second leading cause of cancer-related death. Conventional and advanced treatments currently used for cancer are known for adverse effects and are expensive. Therefore, the search for alternative medicines is necessary. Homeopathy is one of the common complementary and alternative medicine used worldwide for treating and managing various cancers, as it has negligible side effects. However, only a few homeopathic drugs have been validated using various cancer cell lines and animal models. Over the last two decades, an increasing number of validated and reported homeopathic remedies have been developed. Despite the diluted remedies of homeopathic medicine making it controversial clinically, it was found to be more significant as an adjunct therapy for cancer treatment. Hence we aimed to review and summarize the research studies carried out on homeopathic remedies to explore the possible molecular mechanism behind its mode of action against cancer and its effectiveness.

Calcarea carbonica induces apoptosis in cancer cells in p53-dependent manner via an immuno-modulatory circuit.

BACKGROUND: Complementary medicines, including homeopathy, are used by many patients with cancer, usually alongside with conventional treatment. However, the molecular mechanisms underneath the anti-cancer effect, if any, of these medicines have still remained unexplored. To this end we attempted to evaluate the efficacy of calcarea carbonica, a homeopathic medicine, as an anti-cancer agent and to delineate the detail molecular mechanism(s) underlying calcerea carbonica-induced tumor regression. METHODS: To investigate and delineate the underlying mechanisms of calcarea carbonica-induced tumor regression, Trypan blue dye-exclusion test, flow cytometric, Western blot and reverse transcriptase-PCR techniques were employed. Further, siRNA transfections and inhibitor studies were used to validate the involvement of p53 pathway in calcarea carbonica-induced apoptosis in cancer cells. RESULTS: Interestingly, although calcarea carbonica administration to Ehrlich's ascites carcinoma (EAC)- and Sarcoma-180 (S-180)-bearing Swiss albino mice resulted in 30-35% tumor cell apoptosis, it failed to induce any significant cell death in ex vivo conditions. These results prompted us to examine whether calcarea carbonica employs the immuno-modulatory circuit in asserting its anti-tumor effects. Calcarea carbonica prevented tumor-induced loss of effector T cell repertoire, reversed type-2 cytokine bias and attenuated tumor-induced inhibition of T cell proliferation in tumor-bearing host. To confirm the role of immune system in calcarea carbonica-induced cancer cell death, a battery of cancer cells were co-cultured with calcarea carbonica-primed T cells. Our results indicated a "two-step" mechanism of the induction of apoptosis in tumor cells by calcarea carbonica i.e., (1) activation of the immune system of the host; and (2) induction of cancer cell apoptosis via immuno-modulatory circuit in p53-dependent manner by down-regulating Bcl-2:Bax ratio. Bax up-regulation resulted in mitochondrial transmembrane potential loss and cytochrome c release followed by activation of caspase cascade. Knocking out of p53 by RNA-interference inhibited calcarea carbonica-induced apoptosis thereby confirming the contribution of p53. CONCLUSION: These observations delineate the significance of immuno-modulatory circuit during calcarea carbonica-mediated tumor apoptosis. The molecular mechanism identified may serve as a platform for involving calcarea carbonica into immunotherapeutic strategies for effective tumor regression.

Anti-proliferative effects of homeopathic medicines on human kidney, colon and breast cancer cells.

OBJECTIVE: Homeopathy is controversial, due to the claims made for very high dilutions. Although several theories are proposed to understand the mechanisms of action, none are scientifically verified. This study aimed to investigate the efficacy of the selected homeopathic medicines in specific in vitro cancer models. METHODS: We assessed the cytotoxic activity of selected homeopathic medicines in mother tincture (MT), and ultramolecular dilution (30C, 200C, 1M and 10M) against cell lines deriving from tumors of particular organs, Sarsaparilla (Sars) on ACHN cells (human renal adenocarcinoma), Ruta graveolens (Ruta) on COLO-205 (human colorectal carcinoma), and Phytolacca decandra (Phyto) on MCF-7 (human breast carcinoma). Sars was also tested against Madin-Darby canine kidney (MDCK) cells (a non-malignant cell line). Cytotoxicity was measured using the 3-(4, 5-dimethylthiazolyl-2)-2, 5-diphenyltetrazolium bromide (MTT) method, anti-proliferative activity by trypan blue exclusion assay, apoptosis determined by dual staining the cells with ethidium bromide (EB) and acridine orange (AO) dyes. RESULTS: MTs and ultra-diluted preparations of the three homeopathic medicines had highly significant effects in the respective cancer cell lines, producing cytotoxicity and a decrease in cell proliferation. The effects were greatest with the MTs, but in all cases and persisted, although to a lesser degree in the ultra-diluted molecular preparations. Sars showed no effect on MDCK cells. In the homeopathic medicine treated cultures, hallmarks of apoptosis were evident including, cell shrinkage, chromatin condensation and DNA fragmentation. CONCLUSION: This study provides preliminary laboratory evidence indicating the ability of homeopathic medicines as anticancer agents. Further studies of the action of these homeopathic remedies are warranted.

[Antitumor effects of fresh gecko hydrolysate on H22 transplanted tumor in mice].

OBJECTIVE: To compare the inhibitory effects of fresh gecko crude extract and its hydrolysate on H22 transplanted tumor in mice. METHODS: The content of soluble nitrogen (SN-TCA index) was used to determine the degree of enzymolysis. The hydrolysate of gecko was obtained from fresh gecko crude extract by pepsin and papain hydrolyzing. H22 transplanted tumor mouse models were established and divided into negative group, positive group,crude extract group and hydrolysate group. RESULTS: The inhibition rate of the H22 tumor-bearing mice was 29.17%, 48.99% respectively for the crude extract group and the hydrolysate group. The inhibition rate of hydrolysate group and the negative group were significantly different (P < 0.05). The spleen and thymus index for the crude extract group and the hydrolysate group didn't show different compared with the negative group. CONCLUSION: The crude extract of the fresh gecko and the hydrolysate can inhibit the growth of the H22 transplanted tumor. The enzymolysis by pepsin and papain can increase the antitumor activity of the crude extract of fresh gecko.

Marine compounds selectively induce apoptosis in female reproductive cancer cells but not in primary-derived human reproductive granulosa cells.

Anticancer properties of tyrindoleninone and 6-bromoisatin from Dicathais orbita were tested against physiologically normal primary human granulosa cells (HGC) and reproductive cancer cell lines. Tyrindoleninone reduced cancer cell viability with IC50 values of 39 µM (KGN; a tumour-derived granulosa cell line), 39 µM (JAr), and 156 µM (OVCAR-3), compared to 3516 µM in HGC. Apoptosis in HGC's occurred after 4 h at 391 µM tyrindoleninone compared to 20 µM in KGN cells. Differences in apoptosis between HGC and KGN cells were confirmed by TUNEL, with 66 and 31% apoptotic nuclei at 4 h in KGN and HGC, respectively. These marine compounds therefore have potential for development as treatments for female reproductive cancers.

Antihepatoma activity of the acid and neutral components from Ganoderma lucidum.

Ganoderma lucidum extracts or isolated components have been shown previously to acquire many potential biochemical and pharmacological activities, including cancer preventive or antitumor effects. The supercritical fluid extracts of Ganoderma lucidum (total component, TC) and its acid component (AC) and neutral component (NC), were evaluated in vitro and in vivo for their antihepatoma activities. The NC showed a conspicuous inhibitory effect on tumor growth of Heps-bearing mice, whereas AC was less effective. The TC, NC and AC all inhibited the proliferation of BEL-7402 cells through apoptosis pathway and cell cycle arrest. Additionally, the NC and TC induced cell cycle arrest at the G2/M phase, but the AC resulted in a marked increase in the percentage of cells at G1 phase by flow cytometry. It is suggested that NC is an indispensable effective component in terms of antihepatoma activity and its constituents need to be investigated in detail. It was found that the NC, which was detected by GC-MS, contained fatty acids and steroids; hence, it is proposed that some compounds such as long-chain fatty acids and steroids in the NC might also contribute to the antihepatoma activity, although the anticancer activities of G. lucidum traditionally have been considered to be associated with triterpenoids.

[Apoptotic mechanism of gecko crude peptides on human liver carcinoma cell line HepG2].

OBJECTIVE: To investigate the effect of Gecko crude peptides (GCPS) on human liver carcinoma HepG2 cells and its mechanism. METHODS: MTT assay was used to analyze the effect of the GCPS on the proliferation of HepG2 Cell; Nucleus change of HepG2 treated with GCP was observed by Hoechst33258 fluorescence staining, and BAX and BCL-2 were detected with western-blot assay. RESULTS: GCPS could inhibit the proliferation of HepG2 Cell in a time and dosage dependent way, and its half-maximal inhibitory concentration (IC50) was 1.2 mg/mL; HepG2 pretreated with GCPS showed apoptotic morphological changes. GCPS (1.6 mg/mL, 0.8 mg/mL) could decrease the expression of BCL-2 protein, and increase the expression of BAX protein. CONCLUSION: GCPS can inhibit the proliferation of HepG2 cell. The mechanism may be related to the induction apoptosis of HepG2.

[Anticancer activity of Crocodylus siamensis blood in vitro and in vivo].

OBJECTIVE: To study the anticancer effects of the blood of Crocodylus siamensis in vitro and in vivo. METHODS: The inhibitory effects of serum and plasma of Crocodylus siamensis on proliferation of HepG2, BGC823, HeLa and SKOV3 cell were measured by MTT assay. The mouse S180 tumor model was used to evaluate the anti-tumor effect in vivo. RESULTS: High dosage serum and plasma of Crocodylus siamensis could inhibit the proliferation of HepG2, BGC823, HeLa and SKOV3 cell. The tumor inhibitory rate of high dosage blood of Crocodylus siamensis on S180 tumor was up to 57.55%. CONCLUSION: The blood of breeding Crocodylus siamensis has anticancer activity.

[Effect of sodium cantharidinate on the angiogenesis of nude mice with human gastric cancer].

OBJECTIVE: To study the effect of sodium cantharidinate on the angiogenesis of nude mice with human gastric cancer. METHODS: Nude mice xenograft models of human gastric cancer were established by injecting gastric carcinoma cell BGC823 into peritoneal. Expression of VEGF and MVD labeling by CD34 in human gastric cancer cells were measured by immunohistochemistry. RESULTS: Expression scores of VEGF in medium dose and high dose group with sodium cantharidinate treatment were lower than those in low dose and control group (P < 0.01). There was no significant difference between medium dose and high dose group or low dose and control group (P > 0.05). MVD values in medium and high dose group with sodium cantharidinate treatment were lower than those in low dose and control group (P < 0.01), but there was no significant difference between medium dose and high dose group (P > 0.05). CONCLUSIONS: sodium cantharidinate can inhibit the growth of the tumor by down-regulating VEGF expression of the tumour cell and the angiogenesis of the tumour.

[Inhibitory effects of Gecko alcohol extract on human esophageal squamous carcinoma cell line EC-109 proliferation and associated mechanism].

OBJECTIVE: To explore the proliferation inhibition effects of Gecko alcohol extract (GAE) on human esophageal squamous carcinoma cell line EC-109 and its mechanism. METHODS: The inhibitory effects of GAE on proliferation of EC-109 cells were measured by MTT. Nucleolus change of apoptotic cells was observed by Hoechest33342 fluorescence staining. Apoptosis rate of EC-109 cells was detected by flow cytometry. The expressions of apoptosis protein Caspase-3 and FAS in EC-109 cells were investigated by immunohistochemistry. RESULTS: GAE had the inhibition effects on the proliferation of esophageal carcinoma cell EC-109. The apoptosis rate of EC-109 cell treated with GAE(3.0 mg/mL, 4.0 mg/mL) for 48h was 20.63% and 39.73%, respectively. Compared with control group,the expression of Fas and Caspase-3 was significantly up-regulated in GAE treated group. CONCLUSION: GAE can inhibit the proliferation of esophageal carcinoma EC-109 cells and induce them apoptosis which may be correlated with increasing expression of protein Fas and Caspase-3.

[Effect of centipede extract on cervical tumor of mice and its mechanism].

UNLABELLED: To study the anti-tumor activity of centipede extract on cervical tumor of mice and its mechanism. METHODS: The tumor-bearing mice were treated with centipede extract from two solvents [ether (CE) and alcohol (CA)] at different comcentration. The mice' life span, tumor inhibition rate and immune function were estimated. RESULTS: No mice died in CE and CA treatment groups and the tumor inhibition rate was 52.85% and 33.65% respectively. Observed the tumor tissue slices with light microscope and found infiltration of tumor cells in striated muscle in the control group but centipede treatment groups had massive necrosis and apoptosis. Karyopyknosis and apoptotic tumor cells were observed in the treatment groups under transmission electron microscopy. Compared with control group, the expression of Bax increased, the expressions of Bcl-2 and Survivin decreased, but the content of VEGF, the indexes of thymus and spleen had no significant change in treatment groups. The number of CD3+ T lymphocytes had no significant change while the ratio of CD4+ and CD8+, the number of CD19+ B lymphocytes decreaed in the CE group. The numbers of CD3+ and CD4+ lymphocytes decreased in the CA group. The pathological examine indicated no obvious change in the tissue slices of mice's liver and kidney, manifested the concentrations of CE and CA between the article's had no visible side effect. CONNCLUSION: The two extracts (CE and CA) can suppress the growth of cervical tumor and its mechanism may be related to Bax and Caspase-3 medicated the mitochondrial signal transit pathway.

[Effect of cinobufotalin on growth of xenograft of endometrial carcinoma cell line ishikawa in nude mouse and its impact on RRM2 expression].

OBJECTIVE: To investigate the inhibitory effect of cinobufotalin (CBT) on the growth of xenograft endometrial carcinoma cell line ishikawa in nude mice, and its impact on the expression of ribonucleotide reductase subunit M2 (RRM2). METHODS: Eleven nude mice with xenograft were randomly divided into two groups, the CBT group and the control group, which received intra-tumor injection of CBT and saline respectively for one week. The sizes of xenografts were measured before and after the treatment to calculate the inhibition ratio of tumor proliferation; the RRM2-mRNA and protein expressions in tumor tissue were measured by RT-PCR and Western blot respectively. RESULTS: After treatment, the size of xenografts in the CBT group was (0.1314 +/- 0.0304) cm3, which was significantly lower than that in the control group (0.360 0 +/- 0.1145) cm3, (P < 0.05), the tumor proliferation inhibition ratio being 43.46%. The differences of RRM2 mRNA and protein expression levels between the two group were significant (P = 0.019 and P = 0.001). CONCLUSION: CBT significantly inhibits the growth of the xenografts of endometrial carcinoma Ishikawa in nude mice, and the action mechanism is possibly associated with the inhibition on RRM2 expression.