Chemotherapeutic Drug Sensitivity Experiment Of SNK-6 And Hut-78 Cell Lines In Vitro And Multidrug Resistance Gene Detection

Background and ObjectiveT cell lymphoma is a type of malignant proliferative disease which derives from the T lymphocyte with a relative high incidence in Asian countries. Peripheral T cell lymphoma such as nasal type NK/T cell lymphoma and cutaneous T cell lymphoma are more popular than other types with characteristics of high invasion, rapid progression, short survival and poor prognosis, which make these kinds of lymphoma serious threat to the health of our country people. Chemotherapy is a common way to treat lymphoid and hematopoietic malignancies, but there have not been ideal treatments against peripheral T cell lymphoma all over the world at present, and the effectiveness of conventional therapy of the CHOP regimen is poor along with a high relapse rate. Chemotherapeutics have some serious side-effects, low releasing rate, and primary or secondary resistances which will make the treatment fail. Therefore, screening the drug sensitivity of T cell lymphoma which is necessary to find effective treatment methods, and might improve the treatment effectiveness has important clinical significance.Recently, the chemotherapy effectiveness of T cell lymphoma is very poor in clinic, the studies have demonstrated that expression of mdr-1 could be one of the major causes leading to chemotherapy failure. We assumed that there are relationships between the expression of mdr-1 gene and clinical chemotherapy. So we have some further research.In our study, the chemosensitivity of T cell lymphoma cell lines SNK-6 and Hut-78 to antitumor drugs was investigated using by MTT assay in vitro, meanwhile cell inhibition rate and half inhibitory concentration (IC50) were evaluated. And expression of mdr-1 gene of SNK-6 and Hut-78 was detected by RT-PCR to study the relationship between the expression of mdr-1 gene and clinical chemotherapy preliminarily. The results of this study might provide some of theory basis for treatment of T cell lymphoma.Methods1. SNK-6 cells were growing in 1640 with containing 10% human AB type serum and IL-2, Hut-78 were growing in 1640 with containing 10% fetal calf serum, both of them at 37℃in a humidified 5% CO2 incubator. When the cell growth was in exponential phase of growth, tumor cells were collected. The sensitivity of SNK-6 and Hut-78 cells to different concentrations of twelve chemotherapeutic drugs were tested using by MTT assay in vitro, and the IC50 values measured by statistical software.2. The expression of mdr-1 gene mRNA of SNK-6 and Hut-78 cells were detected by Reverse transcription polymerase chain reaction (RT-PCR).Results1. The sensitivity of SNK-6 and Hut-78 cells to cytotoxic drugs varied greatly with different antitumor drugs, and the inhibition rate increased with the increase of the drug concentration in a certain dose range. The sensitivity of chemotherapeutic drugs to SNK-6 from high to low present as follows:gemcitabine, vincristine, docetaxel, etoposide, cisplatin, fluorouracil, doxorubicin, methotrexate, asparaginase, fludarabine, cytarabine, while to Hut-78 as follows:gemcitabine, vincristine, cisplatin, doxorubicin, methotrexate, etoposide, docetaxel, fludarabine, cytarabine, fluorouracil, asparaginase.2. IC50 values were different during two cell lines in vitro treated with the same drug. Specially, both gemcitabine and vincristine were highly sensitive drugs to two cell lines according to the IC50, there were no significant differences (P>0.05) Cisplatin, etoposide, methotrexate and doxorubicin were moderately sensitive drugs, there were no significant differences(P>0.05). Additionly, SNK-6 cell were more sensitive than Hut-78 cell to docetaxel, asparaginase and fluorouracil, there were significant differences (P< 0.05). Compared to other drugs, fludarabine and cytarabine were relatively low/non-sensitive drugs and dexamethasone could not inhibit SNK-6 and Hut-78 effectively in vitro.3. RT-PCR assay showed that mdr-1 mRNA could express in SNK-6 cells and Hut-78 but weaker in latter one.Conclusion1. Different chemotherapeutic drugs have different anti-tumor effect against T cell lymphoma cell lines with high, medium and low sensitivity. The sensitivity of T cell lymphoma cell lines to chemotherapeutic drugs are also different.2. Expression of mdr-1 in SNK-6 and Hut-78 indicate that there are some relevancy between mdr-1 and and clinical chemotherapy efficiency.