ABSTRACT
Objective To investigate the value of methyl thiazolyl tetrazolium assay ( MTT) in predicting drug sensitivity of breast cancer cells in vitro. Methods From January 2010 to July 2016,one hundred and ninety-two patients with breast cancer who underwent modified radical mastectomy or breast conserving surgery (no preoperative radiotherapy or chemotherapy) in the Shanghai Fengxian District Central Hospital were selected. MTT method was used to determine the inhibitory level and sensitivity of 12 drugs and 3 chemotherapy regimens to primary cultured cancer cells of 192 patients with breast cancer. Results (1) The sensitivity of breast cancer cells to 12 drugs were in sequence from high to low as follows: Paclitaxel (PTX)＞ Epirubicin ( EPI )＞ Cisplatin ( DDP )＞ 5-Fluorouracil ( 5-FU )＞ Mitoxantrone ( MIT )＞Vincristine ( VCR )＞ Pirarubicin ( THP )＞ Isosophosphamide ( IFO )＞ Carboplatin ( CBP )＞Cyclophosphamide ( CTX)＞ Methotrexate ( MTX)＞ Changchun Rui bin ( NVB) . The sensitivity of chemotherapy regimens in the three groups from high to low was docetaxel/doxorubicin/cyclophosphamide (TAC )＞cyclophosphamide/epirubicin/fluorouracil ( CEF )＞cyclophosphamide/methotrexate/fluorouracil (CMF). The sensitivity rates of PTX,EPI and DDP were 54%(104/192),42%(81/192) and 37%(71/192) respectively. (2) The average inhibitory rates of DDP,CBP and MIT in stage III breast cancer was higher than those in stage I and II breast cancer,and the differences were statistically significant ( F＝11. 14,4. 303,3. 182,P＜0. 05). (3) HR-breast cancer is more sensitive than HR+breast cancer,PTX, EPI,THP,MIT in HER-2(+) breast cancer is more sensitive than in HER-2(－) breast cancer. Conclusion As a widely used drug sensitivity test method, MTT assay has a certain reference value for screening sensitive drugs and selecting clinical chemotherapy regimens in neoadjuvant chemotherapy of breast cancer. PTX,EPI and DDP are more sensitive to other breast cancer cells than other drugs. Chemotherapy based on in vitro susceptibility results improves the efficiency of chemotherapy and decreases the proportion of changes in chemotherapy schemes due to inefficiency.
ABSTRACT
Objective To investigate whether 5 different chemotherapeutic drugs and their combination of either two drugs could further promote the inhibition on the cell growth of HCC cell line (HepG2) in vitro in the hypoxic and hyponutritional culture medium (HHCM) mimicking the different scenarios of transcatheter arterial chemoembolization (TACE).Methods The cells were treated by 5 drugs for 2 h, 4 h,6 h and 24 h, which include epirubicin (EPI), cisplatin (DDP), mitomycin-C (MMC), oxaliplatin (OXA) and 5-fluorouracil (5-FU) in four concentrations of HHCM (5％, 10％, 25％ and 50％) mimicking the scenarios during TACE and the cell viability was assessed using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay.The combinations of dual drugs treated for 24 h were also tested.Results The sensitive drugs with inhibition rates more than 30％ were EPI, MMC and OXA in 4 different concentrations of HHCM.The sensitivity of the drugs treated for 24 h was significantly increased compared with that for 2 h in 5％, 10％ and 25％ HHCM.The dual combinations did not increase the chemosensitivity of HepG2 cells.Conclusions EPI, MMC and OXA exhibited cytotoxic activity against HepG2 cells in various hypoxia and hyponutrition states.Prolonging the exposure time could increase the sensitivity of drug in HHCM, and the combination of dual drugs cannot enhance the cytotoxic effect.
ABSTRACT
Objective To explore the correlation of chemosensitivity in vitro of anti-cancer drugs between peripheral blood lymphocytes and tumor cells in non-small cell lung cancer of elderly patients.Methods The MTT method was used to test the sensitivity of the peripheral blood lymphocyte and the tumor cells of 52 patients with nonsmall cell lung cancer to 13 kinds of anti-cancer drugs.Results The sensitivity test in eleven drugs include CDDP,CBP,LOHP,PTX,CTX,ICTX,THP,VP-16,GEM,VCR and NVB in the peripheral blood lymphocyte were associated with that in the tumor cells.But no dependablity in two drugs,ADM and HCPT.Conclusion The peripheral blood lymphocytes could be replace tumor cells to test the chemosensitivity of anti-cancer drugs in patients with non-small cell lung cancer.
ABSTRACT
OBJECTIVE: Theoretically, chemotherapy sensitivity and resistance assays help to predict which sensitive agent will be effective for patients. Due to low correlation between in vitro assay results and in vivo responses, chemosensitivity test is not generally applied in actual clinical practices. The aim of this study is to evaluate the influence of cell cycle in the course of cell culture stage on chemosensitivity test as a disturbing factor. METHODS:After synchronization at G0, we conducted experiments on SKOV-3 cell line according to determined cell cycle span (G0, G0/G1, S, G2/M) with MTT (methylthiazolyl-diphenyl- tetrazolium bromide) chemosensitivity test. We evaluated the sensitivity changes of six chemotherapeutic agents (5-FU, Etoposide, Cisplatin, Topotecan, Paclitaxel, Doxorubicin) in each phase at target times. RESULTS: Each phase represented the various results of MTT sensitivity on six chemotherapeutic agents. The variation of sensitivity between experimental (cell cycle synchronized culture) group and reference (conventional culture) group was 21.3+/-5.1 (mean+/-.D)%. CONCLUSION: The cells in the each phase of cell cycle represent different levels of sensitivity to the same chemotherapeutic agent. The required cell culture stage of chemosensitivity test can blur the true candidate agent. This finding can be regarded as one of the reasons of mismatch between in vitro chemosensitivity and in vivo response of candidate chemotherapeutic agents.
Subject(s)
Humans , Cell Culture Techniques , Cell Cycle , Cell Line , Cisplatin , Etoposide , Ovarian Neoplasms , Paclitaxel , TopotecanABSTRACT
OBJECTIVE: Theoretically, chemotherapy sensitivity and resistance assays help to predict which sensitive agent will be effective for patients. Due to low correlation between in vitro assay results and in vivo responses, chemosensitivity test is not generally applied in actual clinical practices. The aim of this study is to evaluate the influence of cell cycle in the course of cell culture stage on chemosensitivity test as a disturbing factor. METHODS:After synchronization at G0, we conducted experiments on SKOV-3 cell line according to determined cell cycle span (G0, G0/G1, S, G2/M) with MTT (methylthiazolyl-diphenyl- tetrazolium bromide) chemosensitivity test. We evaluated the sensitivity changes of six chemotherapeutic agents (5-FU, Etoposide, Cisplatin, Topotecan, Paclitaxel, Doxorubicin) in each phase at target times. RESULTS: Each phase represented the various results of MTT sensitivity on six chemotherapeutic agents. The variation of sensitivity between experimental (cell cycle synchronized culture) group and reference (conventional culture) group was 21.3+/-5.1 (mean+/-.D)%. CONCLUSION: The cells in the each phase of cell cycle represent different levels of sensitivity to the same chemotherapeutic agent. The required cell culture stage of chemosensitivity test can blur the true candidate agent. This finding can be regarded as one of the reasons of mismatch between in vitro chemosensitivity and in vivo response of candidate chemotherapeutic agents.
Subject(s)
Humans , Cell Culture Techniques , Cell Cycle , Cell Line , Cisplatin , Etoposide , Ovarian Neoplasms , Paclitaxel , TopotecanABSTRACT
This paper explains the research status of chemotherapy sensitivity test in three aspects of history of development, specific methods and clinical application. Chemotherapy sensitivity test is an important way to achieve individual treatment of cancer, after more than 60 years, there are two major categories(in vivo method and in vitro method) of more than 10 kinds of methods. The basic steps of sensitivity test inelude culture of primary tumor cells, chemotherapy drugs mixed, the reaction mixture, observing the results of detection and analysis of indicators. This paper focuses on basic principles, main steps and characteristics of six methods, such as the renal capsule of nude mice model of human cancer, the difference between staining cell, tetrazolium salt colorimetric, adenosine triphosphate based bioluminescence tumor chemosensitivity assay, collagen gel embedded culture method and targeting molecule sensitivity assay. Through the results of several clinical trials, it can be seen that chemotherapy under the guidance of drug sensitivity test significantly improved more than experience chemotherapy in efficient rate, median progression-free survival time, survival time, clinical complete remission rate, pathological complete remission rate, etc.
ABSTRACT
Purpose: The adenosine-triphosphate-based chemotherapy response assay (ATP-CRA) is a well-documented and validated technology for individualizing chemotherapy in cancer patients. We evaluate the feasibility of ATP-CRA in colorectal cancer patients. This study will illustrate the assay's success rate, the mean coefficient of variation, and the turnaround time as a validation tool for a chemosensitivity test. Methods: A total of 118 patients, treated by surgery between June 2004 and September 2005 were evaluated for chemosensitivity to seven anticancer agents (5-fluorouracil (5-FU), oxaliplatin, irinotecan, epirubicin, etoposide, gemcitabine, and paclitaxel) by using an ATP-CRA. To allow a comparison between samples, we calculated the chemosensitivity index (CI) based on the percentage cell death rate (CDR, %) at each test drug concentration. Results: The assay success rate was 85.5% (118/138), and the mean coefficient of variation, indicating intra-assay error level, was 9.2%. CDR measured at a therapeutic peak plasma concentration ranged from 0% to 93.6% with a median of 31.0% for 5-FU, 28.5% for oxaliplatin, 34.0% for irinotecan, 25.0% for epirubicin, 21.0% for etoposide, 22.0% for gemcitabine, and 25.2% for paclitaxel. According to the CI, the most sensitive drug varied from patient to patients 10.9% for 5-FU, 10.9% for oxaliplatin, 24.7% for irinotecan, 11.8% for epirubicin, 22.4% for etoposide, 1.1% for gemcitabine, and 23.3% for paclitaxel. Conclusions: Our data suggest that the ATP- CRA is a feasible in-vitro chemosensitivity test in colorectal cancer and that patients show heterogeneous chemosensitivity. A study evaluating the predictive value of ATP-CRA directed therapy is needed to determine the clinical usefulness of the test.
Subject(s)
Mortality , Predictive Value of TestsABSTRACT
Ependymomas usually develop from neuroectodermal organs. Here, we present an ependymoma arising from the pelvic cavity. A 27-year-old Korean female was admitted to the hospital with a sensation of abdominal fullness. Imaging studies revealed a huge heterogeneous nodular mass in the pelvis and lower abdomen. Laparotomy showed that two large masses with multiple nodules were located between the uterus and rectum and uterus and bladder, respectively. Histologically, the tumor was characterized by compact columnar neoplastic cells divided by fibrovascular septae. The neoplastic cells formed true ependymal rosettes and perivascular pseudorosettes. Immunohistochemical staining showed a strong positive reaction for glial fibrillary acidic protein (GFAP) and vimentin and a partial positive reaction for S100 and EMA. The tumor was thus diagnosed as an ependymoma arising from the pelvic cavity. The patient was treated with a debulking operation and chemotherapy based upon the in vitro chemosensitivity test results. The patient was free of cancer for 4 years following surgery. This is a rare case of extraneural ependymoma for which an in vitro chemosensitivity test was critical in determining the multidisciplinary approach for treatment.
Subject(s)
Adult , Female , Humans , Ependymoma/drug therapy , Pelvic Neoplasms/drug therapyABSTRACT
Ependymomas usually develop from neuroectodermal organs. Here, we present an ependymoma arising from the pelvic cavity. A 27-year-old Korean female was admitted to the hospital with a sensation of abdominal fullness. Imaging studies revealed a huge heterogeneous nodular mass in the pelvis and lower abdomen. Laparotomy showed that two large masses with multiple nodules were located between the uterus and rectum and uterus and bladder, respectively. Histologically, the tumor was characterized by compact columnar neoplastic cells divided by fibrovascular septae. The neoplastic cells formed true ependymal rosettes and perivascular pseudorosettes. Immunohistochemical staining showed a strong positive reaction for glial fibrillary acidic protein (GFAP) and vimentin and a partial positive reaction for S100 and EMA. The tumor was thus diagnosed as an ependymoma arising from the pelvic cavity. The patient was treated with a debulking operation and chemotherapy based upon the in vitro chemosensitivity test results. The patient was free of cancer for 4 years following surgery. This is a rare case of extraneural ependymoma for which an in vitro chemosensitivity test was critical in determining the multidisciplinary approach for treatment.
Subject(s)
Adult , Female , Humans , Ependymoma/drug therapy , Pelvic Neoplasms/drug therapyABSTRACT
PURPOSE: A chemosensitivity test can reflect the differences in responses of individual cancer patients to chemotherapeutic agents. The adenosine triphosphate-based chemotherapy response assay (ATP-CRA)is an accurate method, which does not require a large amount of tissue specimen. So far, no studies have evaluated the utility of the ATP-CRA in Korea. Therefore, we investigated the clinical usefulness of the ATP-CRA in 53 patients with lung cancer. MATERIALS AND METHODS: Tumor tissues were obtained from bronchoscopic biopsies or surgical resections. The validity of ATP-CRA was assessed focusing on the success rate, experimental error level (intraassay mean coefficient of variation [CV]) and reproducibility. RESULTS: The overall success rate of ATP-CRA was 90.6% (48/53). Normal cells were effectively eliminated from the tumor tissues with the use of ficoll gradient centrifugation and immunomagnetic separation, which was confirmed using loss of heterozygosity analysis of the 3p deletion. The mean CV of ATP assays was 10.5+/-4.6%. The reproducibility of ATP assays was 94+/-3.8%. The results of the ATP assays were reported to physicians within 7 days of specimen collection. More than 6 anticancer drugs were tested on the tumor specimens obtained from bronchoscopic biopsies. CONCLUSION: The ATP-CRA is a stable, accurate and potentially practical chemosensitivity test in patients with lung cancer.
Subject(s)
Humans , Adenosine Triphosphate , Adenosine , Biopsy , Centrifugation , Drug Therapy , Feasibility Studies , Ficoll , Immunomagnetic Separation , Korea , Loss of Heterozygosity , Lung Neoplasms , Lung , Specimen HandlingABSTRACT
PURPOSE: Cancers are highly individual in their response to chemotherapy, however attempts to predict tumor response to drugs using in vitro cell culture have largely failed. A new technology, the histoculture drug response assay (HDRA), appears to have solved many previous problems. The purpose of this study is to evaluate the reliability of HDRA in a chemosensitivity test for breast cancer. MATERIALS AND METHODS: Tumor specimens from breast cancer patients were evaluated by HDRA using different chemotherapeutic agents. Each specimen was tested using a blind method in order to determine the reproducibility of HDRA results for the same tissue and with a triplicated assay in order to determine reproducibility by different examiners. The evaluative power of this assay and the chemosensitivity of drugs for each specimen was determined. RESULTS: Specimens of 92.9% (65/70) were successfully cultured and evaluated for chemosensitivity. The reproducibility of HDRA for the same tissue was 75% (100% agreement) and 100% (over 70% agreement), respectively. And the reproducibility by different examiners was 78.9% (100% agreement) and 94.7% (over 70% agreement), respectively. Each specimen demonstrated a response to at least one agent. CONCLUSION: The evaluative power and reproducibility of HDRA were high, therefore it might serve as a reliable clinical method for chemosensitivity testing. However, there is a need for clinical trial in which patients are initially randomized for treatment either by HDRA direction or by clinician's choice.
Subject(s)
Humans , Breast Neoplasms , Breast , Cell Culture Techniques , Drug TherapyABSTRACT
PURPOSE: The purpose of the present study is to determine the relation between in vitro resistance to 9 drugs, measured with colorimetric methylthiazol tetrazolium(MTT) assay and prognostic factors. METHODS: Thirty children with leukemia were evaluated at the pediatric department of Dongsan Medical Center. All samples tested with the MTT assay contained 80% or more leukemic cells, which were isolated by Ficoll density gradient centrifugation, and were incubated with 9 drugs for 4 days. The optical density(OD) of the wells was measured with microplate spectrophotometer. Leukemic cell survival(LCS) was calculated by OD treated well/OD control wellsx100(%). LD50 was calculated from the dose-response curve and used as a measure of resistance. RESULTS: Among the 30 children with leukemia, 16 were ALL, 14 were AML. Seventeen boys and thirteen girls ranged in age from 9 months to 16 years. Comparing LD50 values according to leukemic type, AML revealed relatively high LD50 values for all drugs, except VCR. But there were no significant differences between ALL and AML(P>0.05). Male showed high LD50 values for ASP, DET, ARA-C, VP16, ADR and 6TG. Age10 years children showed high LD50 values for all drugs, except 6TG. Patients with a leukocyte count>100,000/mm3 at diagnosis showed high LD50 values for VCR, ASP, DET, MTX, ARA-C, ADR, and 6TG. Patients with normal chromosome showed higher LD50 values. CONCLUSION: Our study showed higher LD50 values at AML, male, ageyears old, leukocyte count>100,000/mm3, and normal karyotype. The MTT test may contribute to the selection of effective chemotherapeutic agent for children with acute leukemia.
Subject(s)
Child , Female , Humans , Male , Centrifugation, Density Gradient , Cytarabine , DEET , Diagnosis , Etoposide , Ficoll , Karyotype , Lethal Dose 50 , Leukemia , Leukocytes , ViperidaeABSTRACT
PURPOSE: Neuroblastoma is the second common solid tumor in chidhood and has the worst prognosis in stage IV case. To improve the future survival rate in this disease the clinical and laboratory characteristics of patients, the characteristics and chemosensitivity test of cultured neuroblastoma cells and the outcome were compared. METHODS: The clinical characteristics including age, urinary catecholamines, serum ferritin, neuron specific enolase, pathology, stage, and patient's outcome were evaluated in four neuroblastoma patients diagnosed at the Department of Pediatrics, Kyungpook National Univesity Hospital. The neuroblastoma cells obtained from tumor or bone marrow cells were cultured and used for immunobead test using mononuclear antibody, biochemical analysis, N-myc oncogene copy, chromosome study and chemosensitivity test. Cyclophosphamide, cisplatin, doxorubicin, etoposide, ifosfamide and melphalan were used in chemosensitivity test. The statistical analyses were done by chi2-method. RESULTS: The age of patients were 6~31 (mean; 18) months. The adrenal gland was the primary site and the stage was IV in all cases. In laboratory data, serum ferritin level were 40~352 (204) ng/mL, neuron specific enolase 45~167 (107) ng/mL, urinary excretion of HVA 1.8~268 (73) mg/day, of VMA 0.2~345 (87) mg/day and the HVA/VMA ratio 0.8~67 (20). Cultured neuroblastoma cells and immunobeads attached around the cell were observed. The partial deletion of short arm or monosomy of chromosome 1, double minutes or homogenous stained region were found in three patients. N-myc oncogene copy was positive in one of two tested. Radical surgery was done in three patients and chemotherpy and radiotherpy by CCG-3881 or -3891 were given in four patients. The duration of survival in three died patients were 6~13 (mean; 10) months. One is survived in relapse-free state for 52 months. In chemosensitivity test, the neuroblastoma cell of survivor was highly sensitive in all drugs compared to the neuroblastoma cell of relapsed patients. CONCLUSION: Normal serum ferritin level or normal chromosome were correlated as a good prognostic factors in survivor compare in relapsed patients. In chemosensitivity test, the neuroblastoma cells of survivor were higher sensitive to all drugs compare to those of relapsed patients. The chemosensitivity test of this method were relatively simple and could be used in selection of anticancer drug and as a prognostic factor in neuroblastoma.
Subject(s)
Humans , Adrenal Glands , Arm , Bone Marrow Cells , Catecholamines , Chromosomes, Human, Pair 1 , Cisplatin , Cyclophosphamide , Doxorubicin , Etoposide , Ferritins , Ifosfamide , Melphalan , Monosomy , Neuroblastoma , Oncogenes , Pathology , Pediatrics , Phosphopyruvate Hydratase , Prognosis , Survival Rate , SurvivorsABSTRACT
Individual tumors, even those of the same histologic type, show varying sensitivity to specific cytotoxic agent. Therefore, sensitivity testing assume an increasingly important as an orientational aid in planning chemotherapy. In the past decade there have been many attempts to develop a chemosensitivity test that would predict the clinical effectiveness of various chemostherapeutic agents against human neoplasms. In the United States National Institue's anticancer drug screening program, a colorimetric assey based on the ability of live cells to reduce a tetrazolium-base compound(MTT) to a blue formazan product was used. There has been an increase in reports of a chemosensitivity assay that use tetrazolium dyes and current the assay is in use in our country. The efficacy of several anticancer drug (vincristine sulfate, Etoposide, doxorubicin CDDP) were evaluated using the in vitro chemosensitivity of MTT assay with two cancer cell lines (MOLT-4, KHOS/NP). The follows obtained. 1) CI50 on MOLT-4 are 0.55ng/ml and 0.81ng/ml for vincristine and oncovin, 142.30ng/ml and 78.75ng/ml for lastet and vepesid, and 19.75ng/ml, 20.43ng/ml and 8.66ng/ml for ADR, ADM and adriblastin, respectively. 2) CI50 on KHOS/NP are 691.35ng/ml, 873.73ng/ml, 1,205.22ng/ml, 768.81ng/ml and 672.19ng/ml for cisplan, cisplatin, cispatin, platinol and cisplatin G, and 9.22ng/ml, 11.46ng/ml and 4.28ng/ml for ADR, ADM and adriblastin, respectively. In conclusion the MTT dye reduction assay to anticancer drug sensitivity using short-term microplate culture might serve as a reliable tool for the selection of effective chemotherapeutic agents in patients with cancers.
Subject(s)
Humans , Cell Line , Cisplatin , Coloring Agents , Doxorubicin , Drug Evaluation, Preclinical , Drug Therapy , Etoposide , United States , VincristineABSTRACT
The MBT-2 mice bladder cancer tissues and the human bladder cancer tissues were implanted on the chorioallantoic membrance(CAM) of the immune deficient fertilized chicken eggs and the histopathologic changes of the CAM and gross morphologic changes of the implanted cancer tissues on CAM ere studied. The chemosensitivity tests using chicken CAM were performed for the 4 human bladder cancer tissues to mitomycin C, thiotepa and adriamycin. With this study, the following results were obtained: 1. The observation of the blood vessel on the chorioallantoic membrane was possible from the post-incubation 6th day group, but for the implantation of the cancer tissues, the blood vessels from the post-incubation 8th day group was appropriate. 2. The budding oif the host capillary vessel to the implanted cancer tissue were observed from the post-implantation second day. 3. The size of the post-implantation 7th day cancer tissues were varied from 2.3 to 9.2 folds to the size of the implantation day. 4. The total failure rate in experiment within post-operative 3rd day were 71.3 percent and the total failure rates in group who had the damage on the chorioallantoic membrance during operation was 82.5 percent. The failure rate of the experiment was declined acutely after post-operative 4th day. 5. The salvage of the eggs could be maintained until post-operative 7th day in 28.1 percent among chemosensitivity test group. 6. The 4 bladder cencer tissues which had the chemosensitivity test showed 1.6 to 7.1 fold growth to the inital implanted size and this meant resistance to the test drugs and these results were corresponded with clinical course.