The Effect And Mechanism Of 2,3-indole Inhibiting Proliferation Of Mouse Melanoma

ObjectiveMalignant melanoma?MM?is one of the most aggressive skin cancers worldwide.It is a cancer derived from melanocytes and has a strong immunog-enicity.The therapeutic effect of malignant melanoma is not ideal.For decades,the treatment of MM mainly relied on chemotherapeutic drugs,especially dacarbazine and its oral analogue,temozolomide,but the efficacy of existing drugs is not stable.This topic has carried out many studies on drug intervention of melanoma in mice,of which 2,3-indole is an endogenous active substance widely present in animals and humans^[1-2],with a clear chemical structure,along with The in-depth study of its pharmacological effects,found that it has a wide range of biological activity,existing studies have demonstrated that ISA has clear analgesic,anti-inflammatory,anti-viral and other effects^[3-6],and for a variety of tumor models The effect is obvious^[7].Therefore,to study the effect of 2,3-indole on the proliferation of mouse melanoma,it is hopeful to provide a new safe drug for the treatment of malignant melanoma.In this study,ISA was used to intervene mouse melanoma in vivo and in vitro to study its effect on the proliferation and apoptosis of melanoma in mice,as well as the mechanism of its potential to reduce the bone marrow suppression of chemotherapeutic drugs.Provides theoretical and experimental evidence for the treatment of malignant melanoma.Methods:1.In vitro experiments:B16 cell lines were divided into four groups:control group,ISA group,positive control group?5-fluorouracil group?,and combination therapy group,each group of six holes.The dosages of each group were as follows:1%DMSO was added to the control group,ISA was added to the ISA group at a final concentration of 400?M dissolved in DMSO?containing 1%DMSO?,and 5-Fu was added to a final concentration of 200?M5-FU?containing 1%DMSO?The combination group was added with two drugs to make the final concentration of 400?M and 200?M?containing 1%DMSO?,respectively,for 48h.Using flow cytometry,constant volume300?l/well,the same speed,fixed collection time 120s/well,cell number detection,cell cycle detection,apoptosis detection,bcl-2 expression detection.2.In vivo experiment:B16 cells were inoculated on the back of C57BL/6mice according to the number of 5*10⁵/branch,and were randomly divided into four groups:control group?gastric administration of western yellow gum?,ISA group?ISA suspension Stomach?,positive control group?5-fluorouracil intragastrically?,combination group?ISA and 5-fluorouracil?.The growth status of the tumors was recorded every day.After two weeks of gavage,the tumors were sacrificed.The weight of the tumor tissues,spleen,bone marrow organs,and other groups were examined for changes in the number of immune cells.Results:1.In vitro results showed that 2,3-indole,5-fluorouracil and their combination have significant effects on inhibiting the proliferation of mouse melanoma cells?P<0.05?.2,3-indole can arrest the cell cycle in S phase,and there is a sub-G1 apoptotic peak?P<0.05?.The positive control 5-fluorouracil has obvious cell cycle arrest,but there is no apoptotic peak,after combined use Apoptotic peaks increased significantly and the cycle arrest effect was more pronounced.Apoptosis results showed that ISA can induce apoptosis of B16cells in a dose-dependent manner?P<0.05?.The results of Bcl-2 showed that the mean fluorescence intensity of Bcl-2 in ISA group was significantly lower than that in the control group.2.In vivo results showed that ISA,5-fluorouracil,and the combination of both can significantly inhibit the growth of mouse melanoma?P<0.05?,and ISA has no obvious inhibition on the immune system,and can even reduce the5-fluorouracil bone marrow suppression?P<0.05?.Conclusion1.2,3-indole has obvious effects on the proliferation of mouse melanoma in vitro and in vivo,indicating that 2,3-indole can effectively inhibit the proliferation of melanoma in mice.The main mechanism may be related to the down regulation of Bcl-2 expression related.2.2,3-indole has no significant effect on the number of normal immune cells,but it can reduce the myelosuppressive effect of 5-fluorouracil and increase the total white blood cells,neutrophils,and NK cell numbers.