Indonesian Journal of Cancer Chemoprevention
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Combination of Doxorubicin and Areca Ethanolic Extract Induces Apoptosis by Increasing Caspase-3 Level on Breast Cancer (T47D) Cells
Get PDFDespite causing many side effects, doxorubicin (Dox) is still one of breast cancer drug of choice. Thus, combination of chemotherapy is developed in order to decrease doxorubicin regimen dose. The aim of this research is to examine the combination effect of doxorubicin (dox) and areca extract (AE) on T47D human breast cancer cells. The cytotoxic activity was determined using MTT assay. The combination index (CI) of the combination treatment was calculated to determine the effects (synergistic, additive or antagonistic). The combination application of dox (6-22nM) and AE (8-30µg/ml) on T47D cells showed synergistic (CI<0.9) or additive effect (CI=0.9-1.1). The effective combination of dox-AE was 6 nM - 8 µg/ml on CI<0.5. Apoptosis induction of AE solely and its combination with dox was then observed using double staining method. Moreover, expression of Bax and caspase-3 protein which mediated apoptosis, were observed using immunocytochemistry. Combination of AE and Dox increased expression of Caspase-3 but did not increase expression of Bax. This result showed AE increase the effectiveness of doxorubicin against T47D cells.Keywords: Breast cancer, doxorubicin, areca extract, T47D cell
Hesperidin Increases Cytotoxic Effect of 5-Fluorouracil on WiDr Cells
Get PDFoai:ojs.ijcc.chemoprev.org:article/63Therapy of colon cancer by using 5-FU often causes problems of resistance. This encourages the development of co-chemotherapy agent. One of the compounds that could potentially be used as a co-chemotherapy agent is hesperidin. This study was conducted to determine the cytotoxic effects of hesperidin, 5-FU and the combination of them, as well as apoptosis induction in colon cancer cells WiDr. Cytotoxic effect of hesperidin, 5-FU, and its combination were observed using MTT assay. Observation of apoptosis was done by double staining method using ethidium bromide-acridin orange. Until 48 hours incubation, hesperidin showed no cytotoxic effects. Cytotoxic effects of 5-FU was observed after 48 hours with the IC50 value of 422 µM. However, hesperidin improved the cytotoxic effects of 5-FU at 48 hour incubation. Either single treatment of hesperidin 200µM or 5-FU 1500 µM did not trigger apoptosis, but combination of them led to the emergence of signs of apoptosis. Based on this study,it can be concluded thathesperidin is potential to be developed as a co-chemotherapy agent of 5-FU on colon cancer but still need further study on its molecular mechanisms.Keywords : hesperidin, 5-fluorouracil, WiDr cells, cytotoxic, apoptosi
Ethanolic Extract of Papaya (Carica papaya) Leaves Improves Blood Cholesterol Profiles and Bone Density in Ovariectomized Rats
Get PDFBone loss and disturbance in the blood cholesterol profiles modulation are two effects caused by menopauses syndromes. As the estrogen concentration in the body decreased drastically, menopause women need the replacement of estrogen to keep the regulation of several physiological functions in the body, such as bone generation and cholesterol regulation in a good condition. Phytoestrogen in Carica papaya leaves, such as quercetin, could be one of the potential agents for the estrogenic effect. The aim of this study is to know the effects of papaya leaf extract (PLE) on the blood cholesterol profiles and bone density in ovariectomized rats. Thirty six female Sprague Dawley rats divided into six groups. The groups were sham-treated ovx (S-OVX), ovariectomized rats (OVX), CMC-Na control (OVX+CMC-Na), positive control (OVX+Estradiol), and the PLE treatment groups dose 750 mg/kgBW (OVX+750mg/kgBW) and dose 1000 mg/kgBW (OVX+1000 mg/kgBW). Administrations of PLE were done in three weeks orally and estradiol administrated intraperitonially. In the end of the treatment, the blood sample of tested animals was collected for the blood cholesterol determination (LDL, HDL, triglyceride, and total cholesterol) and the femur bones were examined for the bone density. Based on the results, PLE dose of 750 mg/kgBW a day in ovariectomized rats showed estrogenic effects in modulating blood cholesterol profile by lowering total cholesterol levels. Meanwhile, PLE dose of 1000 mg/kgBW significantly increased the bone density (p<0.05). Thus, PLE is potential to overcome the negative effects of post-menstrual women especially in the cholesterol blood profiles and bone density.Keywords : Carica papaya, phytoestrogen, bone density, blood cholesterol, ovariectomized rat
The Number of Macrophages and Heterophils on Chick Embryo Chorioallantoic Membrane After Gynura procumbens (Lour) Merr Extract Treatment and bFgF Induction
Get PDFAntiangiogenesis (inhibition of new blood vessels formation) has become a strategy to inhibit cancer development. The aim of this experiment was to investigate antiangiogenic effect of Gynura procumbens (Lour) Merr focusing on the decreasing of the number of macrophages and heterophils on chick embryo chorioallantoic membrane. Nine-days-aged-eggs were divided into six groups (eight eggs each group). Group I (positive control) eggs were induced with bFGF+Tris HCl. Group II (negative control) eggs were treated with DMSO+Tris HCl. Group III (treatment I) eggs were induced with 60 ng bFGF and treated with ethanolic extract of G. procumbens leaves with the dose of 60 µg. The following treatment groups, i.e. group IV (treatment II), group V (treatment III), and group VI (treatment IV) were treated with increasing dose of extract, starting from 75 µg, 90 µg, and the last was 110 µg. Eggs were incubated until they reach the age of twelve days to observe macrophages, while to observe heterophils, eggs were incubated until the age of seventeen days. Based on haematoxylin-eosin staining, macrophages in the treatment groups were less than the control positive group (bFGF+Tris HCl), but based on giemsa staining, the effect of Gynura procumbens in decreasing the number of heterophils could not be observed because some blood smears. These analysis suggest that the ethanolic extract of Gynura procumbens leaves can perform as antiangiogenic agent decreasing the number of macrophages.Keywords: antiangiogenic, macrophages, heterophils, Gynura procumben
Safety Evaluation of Chicken Satay In Yogyakarta Indonesia Based on Benzo[a]pyrene Content
Get PDFChicken satay is one of Indonesian food which is very popular and processed by direct flame over the meat. But, almost every types of food which are prepared by direct flame will produce chemical compounds that contributes to the incidence of cancer. Polycyclic aromatic hydrocarbons (PAH) is a group of compound which can cause cancer and is resulted from the incomplete combustion of fossil fuels or pyrolysis of organic material containing carbon and hydrogen atoms. One PAH compound having mutagenic and highly carcinogenic effect is benzo[a]pyrene which is the highest content found in foods that are burned directly on the flame. This study aims to establish levels of benzo[a]pyrene contained in chicken satay in Yogyakarta, Indonesia and to evaluate how it meets with the maximumbenzo[a]pyrene limit in the burned food based on National Agency of Drug and Food Control which is equal to 5 ppb (5 mg/kg). Analysis were performed by Gas Chromatography method using helium as carrier gas and CPCB-Sil8 as stationary phase. After the sample was prepared and analysed by GC, it is known that the amount of benzo[a]pyrene in chicken satay found in Yogyakarta are ranging from 2.5 to 393.32 ppb. The result are quite high and far above the safety limit determined by National Agency of Drug and Food Control.Therefore, the result of the research needs to be socialized to the public so that people can be more aware about the danger of food they consume.Keywords: chicken satay, benzo[a]pyrene (BaP), policyclic aromatic hidrocarbon (PAH), carsinogen, Yogyakart
The Safety of Areca Seed Ethanolic Extract as Potential Chemopreventive Agent is Proven by Acute Toxicity Test
Get PDFAreca (Areca catechu L.) seeds ethanolic extract (AE) exhibits antiproliferative activity and induces apoptosis on T47D and MCF-7 cells. This study aimed to verify AE safety using acute toxicity test to support its development as chemopreventive agent. Male Sprague Dawley Rat (Rattus norvegicus) age 8 weeks divided into five groups, one group of control treated with 0.5% CMC-Na only and four groups for treatment. Single dose in oral administration was done to test animal with various dose of AE starts from lowest dose to highest dose expected toxic to all of test animal (0.1; 0.72; 5.36 and 10 gram/kgBW). Observation was done during 24 hours and continued for 14 days. The observation criteria were toxic symptoms, appearance and mechanism of toxic effect and pathology of vital organ. Histopathology analysis of some vital organs was done with Haematoxyllin&Eosin (H&E) staining. Toxic effect did not appear either on treatment groups or control group. Treatment of single dose of areca ethanolic extract, even in highest dose, did not cause the death of the animals. Therefore, observation extended to 14 days and terminated by necroption of the animals. All of groups did not show histopathological alterations in microscopic observation. Category of the potential toxicity of AE is practically non-toxic, ie 10 g/kgBW. The result shows the safety of areca seed ethanolic extract which is important for its development as chemopreventive agent.Keywords: Areca catechu, acute toxicity, ra
Antiproliferative Effect of Ethanolic Extract Eugenia uniflora Lam. Leaves on T47D Cells
Get PDFEugenia uniflora Lam. is one of herbal products developed for anticancer. The aim of the present study was to identify the antiproliferative effect of ethanolic extract of Eugenia Uniflora Lam. leaves (EEU) on breast cancer cell line T47D. This Research was initiated by extracting the active contents of Eugenia uniflora Lam. leaves by maceration with ethanol 96%. The extract was then analyzed by thin layer chromatography (TLC). Cytotoxic assay of EEU was carried out by using MTT assay. Apoptosis phenomenon was observed with double staining using acridine orange-ethidium bromide. EEU showed cytotoxic effect on T47D cells with IC50 value of 65 µg/ml. Moreover, EEU 50µg/ml and 100µg/ml induced apoptosis. TLC examination showed that EEU used in this study contain phenolic, flavonoid, and saponin compounds which were suggested to be responsible for antiproliferative effect. Further molecular mechanism underlying EEU antiproliferative effect needs to be done.Keywords: Eugenia uniflora Lam., T47D cells, antiproliferative, apoptosi
Elettaria cardamomum Distillate Increases Cellular Immunity in Doxorubicin Treated Rats
Get PDFDoxorubicin is one of chemotherapeutic agent used for cancer therapy. However, doxorubicin usage causes some adverse effects, such as lymphocyte, CD4+ and CD8+ cells number. Therefore, the co-chemotherapeutic agent is required to reduce the imunosuppression effect. Cardamom (Elettaria cardamomum) contains terpenoid 1,8 cineol. This research aimed to know the effect of Elettaria cardamomum distillate (ECD) in combination with doxorubicin on Sprague Dawley rat’s hematology profile and the amount of CD4+ and CD8+ cells. The experiment was done for 13 days using 6 groups of rats: I doxorubicin (dox) 15 mg/kg BW; II dox + ECD5 mg/kg BW; III dox + ECD 50 mg/kg BW; IV dox + ECD 100 mg/kg BW; V ECD 100 mg/kg BW; VI control without treatment. The hematology profile and the amount of CD4+ and CD8+ were counted before and after treatment using flowcytometer. The results show that ECD increases the amount of lymphocyte, white blood, CD4+ and CD8+ cells in dose dependent manner in doxorubicin treated rats. Based on the data, it can be concluded that ECD is potential to be developed as immunostimulant agent for chemotherapy.Keywords: Cardamom (Elettaria cardamomum), immunostimulant, hematology, CD4+, CD8
Hesperidin Increase Cytotoxic Activity of Doxorubicin on Hela Cell Line Through Cell Cycle Modulation and Apoptotis Induction
Get PDFCombination of chemotherapeutic agent and chemopreventive agent is being a new approach in cancer treatment. This is aimed at enhancing the effectivity and also reducing drug resistance and adverse side effect of the chemotherapeutic agent. Hesperidin, a citrus flavonoid has reported to reduce the proliferation of many cancer cells. The objectives of this study were to investigate cytotoxic activities, cell cycle modulation and apoptosis induction of hesperidin and its combination with doxorubicin on Hela cell lines. MTT [3-(4,5-dimethylthiazol-2-yl)-2.5-diphenyltetrazolium bromide] assay was used to measure the growth inhibitory effect of hesperidin and its combination with doxorubicin on Hela cells. Cell cycle profile was determined by flowcytometry and the data obtained was analyzed by using ModFit LT 3.0 program. Apoptosis assay was done using double staining method using ethidium-bromide and acridine-orange. Hesperidin inhibited cell growth with IC50 48 μM, while the IC50 of doxorubicin was 1000 nM. Combination of 500 nM doxorubicin and 6 μM hesperidin showed strongest inhibitory effect toward Hela cells. Hesperidin of 24 µM accumulated HeLa cells at G1 phase, but its combination with 500 nM Doxorubicin gave G1 and S phase accumulation at 24 h incubation. Both of Hesperidin and Doxorubicin were capable of inducing apoptosis. In accordance of the apoptotic effect, hesperidin, doxorubicin and their combination decreased the expression Bcl-2 and increased the expression of Bax. According to this result, hesperidin has a potency to be developed as co-chemotherapeutic agent for cervical cancer.Keywords: Cochemotherapy, Hesperidin, Doxorubicin, Hela, MTT assa
Lymphocyte Proliferation Activity MTT -Test of Ethanolic Extract of Pasak Bumi Root (Eurycoma longifolia Jack) On Induced 7,12-Dimethylbenz[A] Antracene (DMBA) Female Sprague Dawley Rat
Get PDFPasak bumi root (Eurycoma longifolia Jack) contains quasinoid. Quasinoid potentially has antitumor/anticancer, anti-parasitic, and immunostimulatory activity. The aim of this research was to study the lymphocyte proliferation activity MTT-test of ethanol extract of Eurycoma longifolia Jack root on induced by 7,12-dimethylbenz[a] antracene (DMBA) female Sprague Dawley rat. The test was done at 6 groups of 10 SD rat each. Each groups was administered orally with DMBA (Group I) dose of 20 mg/kg bw, corn oil (Group II), baseline (Group III), and ethanol extract of pasak bumi root dose 12.6; 25.2; and 50.4 mg/kg bw (Group IV,V,and VI). Ethanol extract is given for 5 weeks. At the second week five rats were sacrificed to isolate the lymphocytes of spleen then rats are injected orally with DMBA 2 times a week for 5 weeks. At the eight until twenty fourth week all groups were just administered by aquades. Measurement of spleen relative weight and isolation of the lymphocyte spleen were done at second week and the last of experiment. Lymphocyte proliferation activity were measured by MTT-reduction method. Data were analyzed statistically by analyzes of variance (anova) continued by LSD test and Kruskall Wallis continued by Mann Whitney test. The result showed that limphocyte proliferation activity MTT of ethanol extract of E. longifolia root doses of 12.6; 25.2; and 50.4 mg/kg bw for 14 days prior to DMBA-induced are 339.35 ± 8.43;61.20 ± 6.27;310.28 ± 6.81 and dose 50,4 mg/kg bw after DMBA-induced is 122.108 ± 11.426 .Keyword: E. longifolia root, spleen, lymphocyte cell, 7,12-Dimetilbenz (a) antracene (DMBA