| Purpose:Gliomas are most common in the central nervous system and are primary tumors,accounting for approximately 45%-60% of adult brain tumors.Malignant gliomas show invasive growth,which can easily lead to tumor cell proliferation,recurrence,and migration,making it difficult to completely cure.The treatment is surgical resection,supplemented by postoperative radiotherapy and chemotherapy.With the advancement of modern medicine and the development of technology,more and more means have been gradually applied to the treatment of gliomas.The mechanism of drug treatment of gliomas is not completely understood.Gliomas belong to the scope of brain tumors in Chinese medicine.There are no clear disease names for "brain tumors" in ancient Chinese literature,but there are some similar symptoms in true headache,epilepsy,stroke,dizziness,vomiting,and hiccups.According to the concept of traditional medicine: the overall concept and dialectical treatment,although there is no "brain tumor" disease name,but still can be used to "correct the evil," "adjusting yin and yang," the principle of treatment.The anti-tumor effect of bufalin is a research hotspot in recent years.In this study,the active ingredients extracted from eucalyptus osmanthus were divided into groups to treat glioma cells U87 and U251.Under different time and concentration conditions,different experimental methods were used to detect the effects of bufalin on glioma cells.To explore the mechanism of Qi Ling in treating gliomas,and to provide reference and ideas for future researchers.Material and method:The human glioma cell lines U87 and U251 purchased from the Chinese Academy of Sciences Cell Bank were cultured in vitro.According to previous literature records,the appropriate concentration of bufalin was configured and 6 concentration gradients were applied to the two glioma cells.,Cultured for 24,48,and 72 hours,respectively,using the CCK-8 method to detect the inhibition rate of the glioma cell proliferation and determine the IC50.Based on the IC50,the appropriate concentration of bufalin was calculated.Clone formation assay was used to detect the proliferation inhibitionof glioma cells.Western-blot was used to measure the expression of human telomerase reverse transcriptase(h TERT)protein.Real time PCR was used to detect the expression level of h TERT gene.CCK-8 method was used to detect the growth of human primordial human The killing effect of glioma cells provides a theoretical basis for clinical treatment of gliomas.Results: 1.The concentration of bufalin at the concentration of 5 to 60 nmol/L has a certain inhibitory effect on U251 cells.When the concentration of bufalin is 20 to 160 nmol/L,the proliferation of U87 cells is inhibited in a time and dose dependent manner.The higher the concentration,the longer the duration of action and the more significant the inhibitory effect on the two glioma cells.2.The results of colony formation assay showed that bufalin can effectively inhibit the cloning of U251 and U87 cells in a dose-and time-dependent manner.3.Western-blot results showed that compared with the control group(without bufalin group),bufalin group significantly inhibited the expression of h TERT protein in glioma cells,and the protein expression level decreased with the increase of drug concentration.4.The results of real-time fluorescence quantitative PCR showed that the expression of h TERT-m RNA in U87 and U251 cells of bufalin group was significantly lower than that of the control group(without bufalin group).5.In vitro experiments showed that bufalin has a significant killing effect on human primary glioma cells.Conclusion: 1 Bufalin has the effect of inhibiting proliferation of human glioma cells U87,U251 and human primary glioma cells in vitro and in a dose-and time-dependent manner.2 Bufalin has a significant effect on the expression of human h TERT,inhibits the expression of h TERT,thereby reducing the activity of telomerase and promoting the death of glioma cells. |
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