Shikonin Induction Of Human Glioma Cell Line U251Cells Apoptosis And Related Mechanism Research

Background:Neurospongioma is the most common central nervous system tumors, its tumor tissue from glial cells, poor treatment effect, the recurrence rate is high, high mortality rate. Neurospongioma surgery combined chemotherapy and radiotherapy treatment. Shikonin from comfrey extract of radix arnebiae seu lithospermi root out a anthraquinone compounds. Shikonin derivatives (β,β-acrylic acyl aqua ning, acetyl alkannin main ingredients of radix arnebiae seu lithospermi pigment) is main effective components, alkannin has electron transfer function, absorbed more combined into ester, showed a wide variety of biological activity, promote or interfere with some biochemical reaction process inside the cell, such as anti-inflammatory, antitumor, antibacterial, immune regulation and protect liver, fall blood sugar, anti fertility and antiviral effect. The alkannin could inhibit Neurospongioma and related mechanisms are still unclear, this study chooses U251glioma cell line as the research object, induced by alkannin as drugs, study the effect of shikonin on U251cell apoptosis and related molecular mechanisms.Objective:Explore the shikonin to human glioma cell line U251cells induced apoptosis effect, and its possible mechanism.Method:Choose different concentration of shikonin (final concentration are:1μM、5μM、20μM), acting on U251cells after24h, through CCK8method to test the effects of shikonin on U251cell proliferation, the application of DNA Ladder, Annexin V-FITC/PI double staining to detect cell apoptosis, JC-1after dyeing using fluorescence microscope to observe cells mitochondrial changes, Western blotting-testing Cyt c、AIF、Bax, Bcl2protein expression.Results:Shikonin can inhibit U251cell proliferation, and with the alkannin dose escalation, inhibition enhanced accordingly, maximum inhibition rate of71.20%(71.20±10.32%) DNA Ladder shows:shikonin U251cell, shikonin in each dose group was obviously trapezoidal electrophoresis banding. Flow cytometry instrument testing shows: the shikonin role after U251cells, causing U251cells mitochondrial membrane potential change and leading to cell apoptosis. Cells and with the increase of shikonin dose U251cells apoptosis percentage increase gradually. Western blotting results show that:compared with control group, the shikonin role after U251cells, as the shikonin dose increase with the decreasing concentration of Cyt c、AIF、Bax protein expression increased, while the Bcl-2protein expression decreased.Conclusions:(1) Shikonin can significantly inhibit human glioma cell line U251cells proliferation, has inhibitory effect on cell growth.(2) Shikonin on human glioma cell line U251has a role in inhibition of cell proliferation and apoptosis, performance has certain dose dependent manner.(3) Shikonin induced apoptosis of human U251glioma cell line cells mechanisms, may be related to increase the expression of Cyt c、AIF、Bax protein and cut the Bcl-2protein.