Statistical analysis was carried out by using one way ANOVA followed by Dunnets post hoc test with GraphPad Prism 5.0 (San Diego, CA, USA) and values of P < 0.05 were considered to be statistically significant. RESULTS Preliminary phyto chemical screening The EAAB revealed the presence of various phytoconstituents like alkaloids, glycosides, saponins, triterpenes,
Terpenoids and flavonoids. In vitro cytotoxic study The EAAB has shown concentration dependant cytotoxicity on EAC cells in both Trypan blue [Figure 2] and MTT assays [Table 1], the IC50 values of EAAB were 33.54 ��g and 33.69 ��g respectively in both the assays. Figure 2 In EPZ-6438 molecular weight vitro cytotoxic effect of EAAB on EAC cells by trypan blue exclusion assay. Values are represented as mean �� SEM (n = 3). IC50 for EAAB = 33.54 ��g and 5-FU = 31.39 ��g Table 1 In vitro cytotoxicity of EAAB in EAC cells after 48h exposure by MTT assay In vivo antitumor model Tumor growth response The development of tumor was observed on the day 5, from that day a steady increase in body weight was observed up to end of the study (15th day). The maximum gain of body weight was observed in the EAC control group. In case of EAAB and 5-FU treated groups the body weight was significantly
(P < 0.001) reduced at all the doses [Figure 3]. Figure 3 Effect Selleck Akt inhibitor of EAAB and 5-FU treatment on body weight change in EAC bearing mice. Values are represented as mean �� SEM (n = 6). aP < 0.01, bP < 0.001 as compared to EAC control group The tumor volume, tumor weight and viable cell count were found to be significantly (P < 0.001) decreased and non viable cell count was significantly (P < 0.001) increased in EAAB treated animals at the doses 200 and 400 mg/kg and 5-FU (20 mg/kg) when compared with EAC control animals [Table 2]. Table 2 Effect of EAAB on tumour volume (ml), tumour weight (gm), viable and nonviable cell count (cells��107 cell/ml), mean survival time, percentage increase life span and hematological parameters like RBC (cells��106/��l), WBC (cells��10 ... The EAC bearing
mice administered with EAAB and 5-FU for 14 days and the days of survival were recorded. With EAAB treatment, the survival of EAC bearing mice significantly (P < 0.001) increased as compared to EAC bearing Montelukast Sodium control group. [Figure 4]. Figure 4 Effect of EAAB and 5-FU treatment on median survival time in EAC bearing mice. Data were analyzed using Kaplan�CMeier method. Survival time plotted as days post tumor transplant and ***P < 0.001 when compared to EAC control group In treated group mean survival time was significantly increased to 31.05 �� 2.04 (%ILS = 53.33), 36.12 �� 2.45 (%ILS = 78.37), and 39.78 �� 2.86 (%ILS = 96.44), respectively when compared to EAC control group [Table 2]. Hematological and biochemical parameters There was two-fold increase in the WBC count, drastic fall in the RBC count and hemoglobin content in the EAC control group as compared to normal control group.
Terpenoids and flavonoids. In vitro cytotoxic study The EAAB has shown concentration dependant cytotoxicity on EAC cells in both Trypan blue [Figure 2] and MTT assays [Table 1], the IC50 values of EAAB were 33.54 ��g and 33.69 ��g respectively in both the assays. Figure 2 In EPZ-6438 molecular weight vitro cytotoxic effect of EAAB on EAC cells by trypan blue exclusion assay. Values are represented as mean �� SEM (n = 3). IC50 for EAAB = 33.54 ��g and 5-FU = 31.39 ��g Table 1 In vitro cytotoxicity of EAAB in EAC cells after 48h exposure by MTT assay In vivo antitumor model Tumor growth response The development of tumor was observed on the day 5, from that day a steady increase in body weight was observed up to end of the study (15th day). The maximum gain of body weight was observed in the EAC control group. In case of EAAB and 5-FU treated groups the body weight was significantly
(P < 0.001) reduced at all the doses [Figure 3]. Figure 3 Effect Selleck Akt inhibitor of EAAB and 5-FU treatment on body weight change in EAC bearing mice. Values are represented as mean �� SEM (n = 6). aP < 0.01, bP < 0.001 as compared to EAC control group The tumor volume, tumor weight and viable cell count were found to be significantly (P < 0.001) decreased and non viable cell count was significantly (P < 0.001) increased in EAAB treated animals at the doses 200 and 400 mg/kg and 5-FU (20 mg/kg) when compared with EAC control animals [Table 2]. Table 2 Effect of EAAB on tumour volume (ml), tumour weight (gm), viable and nonviable cell count (cells��107 cell/ml), mean survival time, percentage increase life span and hematological parameters like RBC (cells��106/��l), WBC (cells��10 ... The EAC bearing
mice administered with EAAB and 5-FU for 14 days and the days of survival were recorded. With EAAB treatment, the survival of EAC bearing mice significantly (P < 0.001) increased as compared to EAC bearing Montelukast Sodium control group. [Figure 4]. Figure 4 Effect of EAAB and 5-FU treatment on median survival time in EAC bearing mice. Data were analyzed using Kaplan�CMeier method. Survival time plotted as days post tumor transplant and ***P < 0.001 when compared to EAC control group In treated group mean survival time was significantly increased to 31.05 �� 2.04 (%ILS = 53.33), 36.12 �� 2.45 (%ILS = 78.37), and 39.78 �� 2.86 (%ILS = 96.44), respectively when compared to EAC control group [Table 2]. Hematological and biochemical parameters There was two-fold increase in the WBC count, drastic fall in the RBC count and hemoglobin content in the EAC control group as compared to normal control group.