Antiproliferative Activity and Apoptosis Inducing Mechanism of Anthocephalus cadamba on Dalton's Lymphoma Ascites Cells.

The purpose of this investigation was to evaluate the antiproliferative and apoptogenic mechanistic studies of methanol extract of Anthocephalus cadamba (MEAC) on Dalton's lymphoma ascites (DLA) cells treated mice. Determination of antiproliferative activity was performed by using different DLA cells (2×106 cells, i.p.) inoculated mice groups (n = 12). Groups were treated for 14 consecutive days with MEAC at the doses of 200 and 400 mg/Kg b.w. respectively. The mechanism of antiproliferation activity of MEAC was investigated through morphological studies by acridine orange (AO)/ethidium bromide (EB) double staining method. Comet assay was estimated to check the DNA damage induced apoptosis property. Furthermore, flow cytometry (FACS) was used to quantitatively detect the apoptotic rate by double labeling techniques using Annexin-V FITC/propidium iodide staining analysis and apoptotic proteins expression done by western blotting assay method. MEAC exhibited significant (p<0.01) decrease the tumor volume, viable cell count, tumor weight and elevated the life span of DLA tumor bearing mice. Analysis of AO/EB staining and flow cytometry showed that MEAC possessed apoptosis induced antitumor activity on DLA cells in a dose dependant manner. Dose dependent induction of DNA damage on DLA cells were observed after MEAC treatment, which was evident from the appearance of comet tail length. Pro-apoptotic gene, Bax was up-regulated and down-regulation of the Bcl-2/Bax ratio, suggesting that Bcl-2 family involved in the control of apoptosis. Experimental results revealed that MEAC possess potent antitumor activity via induction of cancer cell apoptosis mechanism.

Methanolic extract of Anthocephalus cadamba induces apoptosis in Ehrlich ascites carcinoma cells in experimental mice.

OBJECTIVE: Anthocephalus cadamba (Roxb.) Miq. (Family: Rubiaceae), a folk medicine commonly known as "Kadam" in Bengali, has been used for the treatment of tumor. The methanolic extract of A. cadamba (MEAC) showing antitumor activity on Ehrlich ascites carcinoma (EAC) cells treated mice was already reported. This study was designed to study the apoptosis-inducing property of MEAC and its mechanism in EAC cells in mice. MATERIALS AND METHODS: Apoptogenic morphology was determined by fluorescent DNA-binding double staining method using dyes acridine orange (AO)/ethidium bromide (EB). Comet assay was estimated to check the DNA damage. Flow cytometry (fluorescence-activated cell sorting [FACS]) was used to detect the apoptotic rate quantitatively by double labeling techniques using annexin V FITC/propidium iodide staining. Apoptotic protein expression was done using Western blotting assay method. STATISTICAL ANALYSIS: Results are expressed as mean ± standard deviation. Statistical analysis was performed using ANOVA followed by Dunnett's post hoc test of GraphPad Prism software. *P < 0.05, **P < 0.01 and ***P < 0.001 were considered statistically significant. RESULTS: Apoptosis-inducing effect of MEAC on EAC cells was confirmed from AO/EB staining and FACS analysis. MEAC treatment showed dose-dependent induction of DNA damage. Apoptosis was induced by increasing the expression of multiple downstream factors such as pro-apoptotic protein p53 and p21 in EAC. Bax was up-regulated and anti-apoptotic protein Bcl-2 was down-regulated resulting in decrease of the Bcl-2/Bax ratio by MEAC treatment. CONCLUSION: Experimental results revealed that MEAC induces apoptosis by modulating the expression of some pro-apoptotic and anti-apoptotic proteins in EAC and thus exerts its anti-tumor activity.

Apoptogenic effects of ß-sitosterol glucoside from Castanopsis indica leaves.

ß-Sitosterol glucoside (BSSG) is a natural biologically active substance isolated from the Castanopsis indica leaves. This study explored the apoptogenic mechanistic studies of BSSG against Ehrlich's ascites carcinoma (EAC) treated mice through morphological study, comet assay, flow cytometry (FACS) and Western blotting assay method. AO/EB staining and FACS analysis showed that BSSG possessed apoptosis induction activities on EAC cells. Dose dependent induction of DNA damage was observed after BSSG treatment. Increase the expression of apoptotic protein p53 and p21 in EAC, multiple downstream factors contributing to apoptosis pathway. The increase of caspase-9 and caspase-3 activities revealed that caspase was a key mediator of the apoptotic pathway induced by BSSG, and up-regulation of Bax and down-regulation of anti-apoptotic protein Bcl-2 resulted in the decrease of Bcl-2/Bax ratio. Owing to the combination of significant antitumour activity by inducing apoptosis, BSSG holds the promise of being an interesting chemo-preventive agent active in cancer therapy.

Antitumor activity and antioxidant status of Streblus asper bark against Dalton's ascitic lymphoma in mice.

Streblus asper Lour (Moraceae), commonly known as Siamee Rough Brush in English is widely distributed in subtropical Asia and traditionally used for several medicinal purposes. In the present study, the ethyl acetate fraction of the methanol extract from Streblus asper bark (EASA) was evaluated for antitumor effect against Dalton's ascitic lymphoma (DAL) in Swiss albino mice. Twenty-four hours after intraperitoneal inoculation of DAL cells in mice, EASA was administered intraperitoneally at 200 and 400 mg/kg body weight for 9 consecutive days. On the 10th day, half of the mice were sacrificed to determine the tumor growth parameters, and the rest were kept alive for survival assessment. Hematological, serum biochemical and tissue (liver, kidney) antioxidant profiles were also determined. EASA exhibited significant and dose dependent decrease in tumor growth parameters and increased survival of DAL bearing animals. EASA significantly and dose-dependently normalized the altered hematological, serum biochemical and tissue antioxidant parameters as compared with the DAL control mice. From the present study it may be concluded that S. asper bark possesses remarkable antitumor efficacy mediated by amelioration of oxidative stress by multiple mechanisms.

Cat's whiskers flavonoid attenuated oxidative DNA damage and acute inflammation: its importance in lymphocytes of patients with rheumatoid arthritis.

Cleome gynandra L. (Capparidaceae) is one of the vegetables commonly known as 'Hurhur' and 'Karaila' in India, 'Pe Hua Tsai' in China and "Cat's whiskers" in English. Present study was aimed to characterize previously isolated Cat's whiskers flavonoid as 5-hydroxy-3, 7, 4' -trimethoxyflavone (5HTMF) and to evaluate its effect on carrageenan-induced acute inflammation in rats and hydrogen peroxide induced DNA damage in mouse macrophages. The ex vivo effect of 5HTMF upon generation of free radicals in the mononuclear lymphocytes of patients with rheumatoid arthritis (RA) was also evaluated. 5HTMF not only reduce the swelling of hind paw in rats from 1 to 3 h of carrageenan injection but also decreased serum nitric oxide (NO) production. Toxic hydrogen peroxide induced oxidative DNA damage that was significantly decreased by 5HTMF. Though oxidative stress is a potential biomarker for determining disease activity in patients with RA, surprisingly 5HTMF inhibited the superoxide, hydroxyl and NO radicals in the isolated peripheral blood mononuclear lymphocytes of patients with RA. From the above study, it may be concluded 5HTMF attenuated acute inflammation by inhibiting NO and by protecting the oxidative DNA damage due to hydrogen peroxide scavenging property. It was also equally effective in scavenging the free radicals in lymphocytes of patients with RA. Collectively, our results indicate that 5HTMF as well as leafy vegetable of Cat's whiskers may be a promising nontoxic food alternative in attenuating the oxidative stress, meriting further studies on other human inflammatory cells.

Antitumor activity and antioxidant property of Curcuma caesia against Ehrlich's ascites carcinoma bearing mice.

CONTEXT: Curcuma caesia Roxb. (Zingiberaceae), commonly known as "Kala Haldi" in Bengali, has been traditionally used for the treatment of cancer, bruises, inflammation and as an aphrodisiac. OBJECTIVE: To evaluate the antitumor activity and antioxidant status of the methanol extract of Curcuma caesia (MECC) rhizomes on Ehrlich's ascites carcinoma (EAC)-treated mice. MATERIALS AND METHODS: In vitro cytotoxicity assay of MECC was evaluated by using Trypan blue method. Determination of in vivo antitumor activity was performed after 24 h of EAC cells (2 × 10(6) cells/mouse) inoculation; MECC (50 and 100 mg/kg i.p.) was administered daily for nine consecutive days. On day 10, half of the mice were sacrificed and the rest were kept alive for assessment of increase in lifespan. Antitumor effect of MECC was assessed by the study of tumor volume, tumor weight, viable and non-viable cell count, hematological parameters and biochemical estimations. Furthermore, antioxidant parameters were assayed by estimating liver and kidney tissue enzymes. RESULTS: MECC showed direct cytotoxicity (IC50 90.70 ± 8.37 µg/mL) on EAC cell line. MECC exhibited significant (p < 0.01) decrease in tumor volume, tumor weight, viable cell count and percentage increased the lifespan (57.14 and 88.09%) of EAC-treated mice. Hematological profile, biochemical estimation, tissue antioxidant assay significantly (p < 0.01) reverted to normal level in MECC-treated mice. CONCLUSION: MECC possesses potent antitumor activity that may be due to its direct cytotoxic effect or antioxidant properties. Further research is in progress to find out the active principle(s) of MECC for its antitumor activity.

Antitumor activity and antioxidant role of Streblus asper bark against Ehrlich ascites carcinoma in Swiss albino mice.

The present study assessed the methanol extract of Streblus asper stem bark (MESA) for antitumor effect and antioxidant influence against Ehrlich ascites carcinoma (EAC) in Swiss albino mice. Twenty four hours after intraperitonial inoculation of tumor (EAC) cells in mice, MESA was administered at 200 and 400 mg/kg body weight daily for 9 consecutive days. On the 10 th day, half of the mice were sacrificed for estimation of tumor parameters, haematological, liver and kidney antioxidant parameters; and the rest were kept alive for assessment of survival. MESA exhibited dose dependent and significant (p < 0.01) decrease in tumor proliferation and extended the life span of EAC bearing mice. Hematological profiles were significantly (p < 0.01) restored near to normal in MESA treated mice as compared to EAC control. MESA treatment significantly (p < 0.01) modulated the hepatic and renal antioxidant parameters as compared to EAC control. The present study demonstrated that S. asper bark possessed promising antitumor efficacy in mice, plausibly mediated by amelioration of oxidative stress by multiple mechanisms.

Evaluation of antitumor activity of Mimusops elengi leaves on Ehrlich's ascites carcinoma-treated mice.

CONTEXT: Mimusops elengi (M. elengi) Linn. (Sapotaceae) has been used as a folk medicine in wound healing, and the treatment of pain, and inflammation in many parts of India. OBJECTIVE: The purpose of this investigation was to explore the antitumor activity of methanol extract of M. elengi (MEME) in Swiss albino mice against Ehrlich ascites carcinoma (EAC) cell line. MATERIALS AND METHODS: Twenty-four hours after intraperitoneal (i.p.) inoculation of tumor (EAC) cells in mice (n = 12), MEME was administered at 200 and 300 mg/kg body weight daily for 9 consecutive days. On day 10, half of the mice were dissected and the rest were kept alive for assessment of increase in life span. The antitumor effect of MEME was assessed by evaluating tumor volume, viable and nonviable tumor cell count, tumor weight, hematological parameter, and biochemical estimations. In vivo antioxidant parameters were assayed by estimating liver tissue enzyme. In vitro cytotoxicity assay of MEME was measured by using trypan blue exclusion method. RESULTS AND DISCUSSION: MEME showed significant (p < .001) decrease in tumor volume, packed cell volume, and viable cell count, and increased the life span of EAC bearing mice. Hematological, biochemical profile, and in vivo antioxidant parameters were significantly restored toward normal levels in MEME-treated mice as compared to EAC control. MEME also showed direct cytotoxicity on EAC cell line in a dose-dependent manner. CONCLUSIONS: The present study demonstrates that M. elengi leaves exhibited antitumor activity in Swiss mice, which may be due to its cytotoxic effect and antioxidant properties.

Antitumor potential of Castanopsis indica (Roxb. ex Lindl.) A. DC. leaf extract against Ehrlich's ascites carcinoma cell.

Methanol extract of C. indica (MECI) leaves showed direct cytotoxicity on Ehrlich ascites carcinoma (EAC) cell in a dose dependant manner and there was significant decrease in the tumor volume, viable cell count, tumor weight and elevated the life span of EAC tumor bearing mice. Hematological profile and biochemical estimations were significantly restored to normal levels in MECI treated as compared to EAC control mice. MECI treatment significantly modulated the tissue antioxidant assay parameters as compared to the EAC control mice. The results revealed that MECI possesses significant dose dependent antitumor potential which may be due to its cytotoxicity and antioxidant properties.

Evaluation of antitumor activity and in vivo antioxidant status of Anthocephalus cadamba on Ehrlich ascites carcinoma treated mice.

ETHNOPHARMACOLOGICAL RELEVANCE: Anthocephalus cadamba (Roxb.) Miq. (Family: Rubiaceae) is commonly known as "Kadamba" in Sanskrit and Hindi in India. Various parts of this plant have been used as a folk medicine for the treatment of tumor, wound healing, inflammation and as a hypoglycemic agent. AIM OF STUDY: The purpose of this investigation was to evaluate the antitumor activity and antioxidant status of defatted methanol extract of A. cadamba (MEAC) on Ehrlich ascites carcinoma (EAC) treated mice. MATERIALS AND METHODS: In vitro cytotoxicity assay has been evaluated by using the trypan blue method. The determination of in vivo antitumor activity was performed by using different EAC cells (2 × 10(6) cells, i.p.) inoculated mice groups (n=12). The groups were treated for 9 consecutive days with MEAC at the doses of 200 and 400 mg/kg b.w. respectively. After 24h of last dose and 18 h of fasting, half of the mice were sacrificed and the rest were kept alive for assessment of increase in life span. The antitumor potential of MEAC was assessed by evaluating tumor volume, viable and nonviable tumor cell count, tumor weight, hematological parameters and biochemical estimations. Furthermore, antioxidant parameters were assayed by estimating liver and kidney tissue enzymes. RESULTS: MEAC showed direct cytotoxicity on EAC cell line in a dose dependant manner. MEAC exhibited significant (P<0.01) decrease in the tumor volume, viable cell count, tumor weight and elevated the life span of EAC tumor bearing mice. The hematological profile, biochemical estimations and tissue antioxidant assay were reverted to normal level in MEAC treated mice. CONCLUSION: Experimental results revealed that MEAC possesses potent antitumor and antioxidant properties. Further research is going on to find out the active principle(s) of MEAC for better understanding of mechanism of its antitumor and antioxidant activity.