In vitro and in vivo evaluation of a standardized Curcuma longa Linn formulation in cervical cancer.

BACKGROUND: The anti-cancer activity of phytomolecules present in turmeric or haridra (Curcuma longa Linn) extracts against cancer has been described in various 'in vitro and in vivo' studies. OBJECTIVE: In the present study, in vitro and in vivo anti-cancer and chemo-preventive activity of a new standardized Supercritical Turmeric Oil Extract (SCTOE) NBFR-03 was evaluated in cervical cancer models. METHODS AND MATERIALS: In vitro cytotoxicity of this formulation was assessed at 10, 20, 40, and 80 µg/ml concentrations, in three cervical cancer cell lines (HeLa, SiHa, ME180) using Sulforhodamine B assay. The in vivo anti-cancer activity was evaluated in two groups of female nude mice; the first one was with tumor xenograft implants and at the same time treatment was started with 96 µl/kg/day p.o. and 192 µl/kg/day p.o. NBFR-03 for three months. The second group was kept as chemoprevention group where mice were pre-treated with the formulation (96 µl/kg/day p.o.) for two weeks and injected with cancer cell suspension with continued treatment for three months. RESULTS: No cytotoxicity was seen in any cell line with the extract when compared to positive control (Adriamycin 10 µg/ml). In mice the first treatment group with tumor xenograft implants did not show any significant anti-tumor activity but showed a trend where higher dose group had smaller tumor volumes as compared to lower dose group and controls (p = 0.37 and p = 0.34 respectively). The chemopreventive group with pre-treated mice also showed smaller tumor size as compared to controls (p = 0.163). CONCLUSION: NBFR-03 turmeric oil extract showed a promising trend in mice pre-treated with NBFR-03. There is a scope for further studying the potential of this extract as complementary therapy and as a chemopreventive.

In vitro chemosensitivity profile of oral squamous cell cancer and its correlation with clinical response to chemotherapy.

CONTEXT: Oral cancers represent a disparate group of tumors with diverse clinical behavior and chemosensitivity profile. Currently, it is difficult to predict whether a tumor will respond to chemotherapy and which drug(s) will achieve the maximum clinical response. AIMS: To study in vitro chemosensitivity profile of oral cancers and to correlate the in vitro chemosensitivity of oral cancer to clinical response to chemotherapy. SETTINGS AND DESIGN: Prospective study in a tertiary cancer care center. METHODS AND MATERIAL: We prospectively studied the chemosensitivity profile of 57 untreated, advanced, unresectable oral cancers to cisplatin, methotrexate, 5-fluorouracil and their combinations by using histoculture drug response assay (HDRA) and correlated them to the clinical response to chemotherapy. STATISTICAL ANALYSIS USED: Chi Square test. RESULTS: Biopsy samples were successfully histocultured in 52/57 (91%) cases. Of these 52 evaluable patients, 47 had primary gingivo-buccal cancers and five had tongue / floor of mouth cancers. Based on the assay, 27 (52%) tumors were sensitive to cisplatin, 27 (52%) to methotrexate, 24 (46%) to 5-fluorouracil, 38 (73%) to combination of cisplatin and methotrexate and 36 (69%) to combination of cisplatin and 5-fluorouracil. Of these, 31 patients with good performance status received two cycles of chemotherapy using one or more of these test drugs. There was a significant correlation (p=0.03) between the in vitro chemosensitivity and the clinical response. Negative predictive value of the test was 80%, positive predictive value-69%, sensitivity-79% and specificity -71%. The overall accuracy of the assay was 74%. CONCLUSIONS: We found HDRA to be a fairly good predictor of chemo-response of oral cancer.

Effect of cisplatin-based chemotherapy on emergence of cisplatin resistance, and its correlation with intracellular glutathione levels and accumulation of p53 protein in human ovarian cancer.

Forty-seven ovarian cancer cases in which 20 were previously treated with cisplatin (cisPt) based chemotherapy, were checked for in vitro chemosensitivity using MTT assay. The drugs included in the study were cisPt, adriamycin (ADR), epirubicin (EPR) and etoposide (ETO). The logarithemic concentrations (0.1, 1.0, 10.0 and 100.0 micrograms/ml) of these drugs were used in the MTT assay. The IC50 values for these drugs in the above tumor samples were calculated. The effect of pretreatment with cisPt based chemotherapy on the emergence of drug resistance, expression of p53 protein (detected using immunohistochemical method by employing monoclonal antibody to p53) and intracellular glutathione (GSH) levels was also studied. Our results demonstrated the superiority of EPR in terms of its efficacy as compared to the other drugs used in the study. EPR was effective in both, previously cisPt-exposed and cisPt-unexposed ovarian cancer cases indicating its importance as a second line chemotherapy in the refractory ovarian carcinoma cases. Pre-exposure to cisPt based chemotherapy appears to result in the emergence of cisPt resistance, elevated intracellular GSH levels as well as p53 positivity. A statistically significant correlation was also observed between ADR and EPR resistance and p53 positivity (P < 0.01 and 0.05 respectively).

Chemosensitivity of advanced larynx carcinoma cells in vitro and significance of multidrug resistance markers in these tumors.

Thirty cases of previously untreated advanced larynx carcinoma were checked for in vitro chemosensitivity and presence of the resistance markers viz. P-glycoprotein (P-gp) glutathione-S-transferase-pi (GST-pi) and protein kinase C (PKC) overexpression. The cytotoxicity testing was done using MTT assay and the resistance markers were checked by immunohistochemical methods using monoclonal antibodies. The drug combinations employed in MIT assay were 5FU* + MTX*, 5FU + cisPt*, 5FU + Mito*, cisPt + Mito and MTX + Mito (*5FU = 5Fluorouracil, MTX-methotrexate, cisPt-cisplatin and Mito = mitomycin C). No statistically significant correlation was observed between resistance to the above drug combinations and presence of the resistance markers under consideration. A statistically significant correlation was observed between node positivity and expression of resistance markers which indicates that presence of one or more of these markers in these tumors may be considered as a negative prognosis marker. CisPt-Mito was found to be the most effective drug combination in vitro, in the cases studied.

Circumvention of drug resistance of P388/R cells by the combination of adriamycin and mitoxantrone with hyperthermia (42 degrees C).

P388/R, the adriamycin (ADR) resistant subline of murine P388 lymphocytic leukemia was cross-resistant to the drug mitoxantrone (MTN). One hour hyperthermia at 42 degrees C (HT) was employed along with ADR (10 micrograms/ml) and MTN (10 micrograms/ml) for circumventing the drug resistance of P388/R cells in in vitro-in vivo bioassays. Inhibition in the incorporation of tritiated thymidine into cellular DNA was measured to check the in vitro cytotoxic effect. Hyperthermia, ADR and MTN alone could not bring about significant degree of inhibition of DNA biosynthesis whereas the combination of ADR and MTN along with HT resulted in a synergistic cytotoxic action (p less than 0.001 and 0.01, respectively). The cells were treated with the drugs in vitro and inoculated into BDF1 mice. It was observed that ADR, MTN or HT pretreatment of the cells resulted in an increase of the life-span of the mice by 4.0-25.0%, 10-20% and 44-50%, respectively, whereas the pretreatment of cells with the combination of ADR and MTN with HT resulted in an increase by 104-125% and 212-220% in life-span of the mice, respectively. Studies revealed that the combination ADR-HT and MTN-HT resulted in circumvention of resistance of P388/R cells to ADR and MTN.

In vitro modulation of adriamycin and mitoxantrone cytotoxicity by hyperthermia and diazepam, in human chronic myeloid leukemia cells.

Adriamycin and mitoxantrone are known antitumor agents. The use of these agents is limited by their toxicity to normal body tissue. This paper shows that it is possible to achieve greater log cell-kill by using these drugs in combination with hyperthermia and diazepam. Experiments were carried out on 22 human chronic myeloid leukemia samples. 10 micrograms/ml adriamycin and 1 microgram/ml mitoxantrone were used in combination with hyperthermia 42 degrees C, for 3 h and 1 h respectively, with and without diazepam (1 microgram/ml). Inhibition of incorporation of radiolabeled nucleic acid precursor (3H-thymidine) in treated cells as compared to the untreated cells was used as a measure of cytotoxicity. The statistical evaluation of the data shows that the enhancement of drug cytotoxicity due to hyperthermia and diazepam is highly significant (p less than 0.001) in case of both the drugs.

Inhibition of DNA biosynthesis by vincristine and pentoxifylline in murine P388 leukemia cells resistant to doxorubicin.

Pentoxifylline (PTX) is a methylxanthine used clinically in the treatment of intermittent claudication. It is an active hemorheological agent used for the treatment of defective microcirculation. The use of the anticancer agent vincristine is limited by its toxicity to normal body tissues. The data presented in the present paper show that it is possible to achieve greater cell-kill by using vincristine in combination with pentoxifylline. The effect of pentoxifylline alone and in combination with vincristine was studied using membrane filtration technique in P388 leukemia (P388) and its subline P388/DOX resistant to doxorubicin and cross-resistant to vincristine. Pentoxifylline (100 mumol/l) had minimal inhibitory effect on DNA biosynthesis in P388 leukemia cells. Vincristine, at the concentration employed in this study did not show significant inhibition of DNA biosynthesis confirming multidrug resistant nature of P388/DOX cells. Pentoxifylline had a dose-sparing effect, wherein it enhanced the antiproliferative activity of vincristine at a clinically achievable concentration. The studies on reversibility of inhibition of DNA biosynthesis in P388/DOX cells pretreated with vincristine and pentoxifylline showed the irreversible nature of the effect of combination of vincristine and pentoxifylline. This observation warrants the possible use of pentoxifylline as an adjuvant in cancer chemotherapy.

Effect of mitoxantrone on human chronic myeloid leukemia cells in vitro, combined with hyperthermia.

Mitoxantrone, a new anticancer drug has DNA-binding properties similar to anthracycline antibiotics. In the present studies, effect of the drug has been tested in vitro on human chronic myeloid leukemia cells at 37 degrees C and 42 degrees C. Inhibition of 3H-tritiated thymidine incorporation in the drug-treated cells compared with untreated cells has been used as the parameter of cytotoxicity of the drug and hyperthermia. Cell samples from 11 CML patients who did not receive any chemotherapy showed less response to the drug at 0.5 micrograms/ml and 1 microgram/ml at 37 degrees C. Exposure of CML cells to 42 degrees C for 2 h indicated 13 to 44% inhibition in 3H-TdR incorporation. However, when CML cells were exposed to mitoxantrone for 2 h at 42 degrees C the 3H-thymidine incorporation was inhibited to the extent of 27 to 71%, indicating greater cellular damage with this combination.

Chloramphenicol: screening and review to evaluate its potential beneficial effects in leukaemia.

Chloramphenicol is an antimicrobial agent having a very broad-spectrum of activity including Gram-positive bacteria, Gram-negative bacteria and anaerobes. However the use of chloramphenicol has reduced over a period of time due to the adverse effects of causing bone marrow depression or in some cases severe aplastic anaemia. As the effects are seen on the bone marrow cell, it was intended to find out if these adverse effects could be used for the benefits in leukaemia patients, using in-vitro study on leukaemic cell lines. The study showed inhibition of growth of the leukaemia cells by chloramphenicol which was comparable to or better than daunorubicin in some cell lines. The article also discusses the other adverse effect profile of chloramphenicol compared with anticancer drugs and its potential benefit in leukaemia and in neutropenic fever.