Experiment Study On The Effect Of Oxymatrine In Bladder Cancer Cell Lines T24 In Vitro

Background and objective:Bladder cancer is the most common genitourinary neoplasm in China,which has a high recurrence rate with intravesical chemotherapy following TURBT and partial cystectomy. Tradition Chinese medicine has unique advantage in treating many diseases. There are few reports of prevention and cure on bladder cancer. Some new researches showed that oxymatrine (OMT) could suppress the proliferation of some cancer cells as galactophore cancer, lung cancer and gastro cancer in vitro. To study the effects of OMT on biology characters of T24 bladder cancer cells, and its associated mechanisms of apoptosis, we treated T24 cells with OMT by several techniques.Methods: 1. T24 cells were treated with OMT in vitro. The cell inhibition rate was measured by MTT assay. Morphologic changes were studied with optic microscope and transimission electron microscope (TEM). Distribution of cells cycle and cells apoptosis were determined by flow cytometry(FCM). 2. The expression of oncoprotein and anti-oncogene protein P53, Bcl-2, Bax and Caspase-3 were determined by immuno- cytochemity in our reseach. Results: 1. Anti-proliferation rate of T24cells by OMT treated were 13.75～91.03％, for 72h at 0.625mg/ml~10mg/ml, in a time and dose dependent way. OMT inhibited T24 cells growth with IC50 1.82mg/ml. Treated with 1.25mg/ml OMT for 72h, some T24 cells condensed with membrane buding, the nuclear chromatin compaction and membrane-closed apopototic bodies formation by optic microscope and TEM. T24 cells group G0/G1 were 43.31%,S 54.10%,G2/M 2.59% respectively by FCM； Exposed 1.25mg/ml OMT for 72h, they were 54.52%, 44.42%,1.06% , respectively and apoptosis cells were 5.37%；Exposed 2.5mg/ml OMT for 72h, they were 52.41%,40.59%,6.99% , respectively.2. The protein expressions of P53, Bcl-2, Bax Caspase-3 in T24 cells control group were 76.25%, 64.38%, 27.38%, 18.13% respectively；T24 cells group treated with 1.25mg/ml OMT for 72h, 25.75%,27.13%, 58.25%, 38.5% respectively. There were statistically significance between control group and OMT treaed gruops (p < 0.05).Conclusions: 1. Proliferation of T24 cells were suppressed statistically by 1.25mg/ml OMT. Anti-proliferation was 91.03%, when T24 cells were expesoed OMT for 72h. Morphologic changes of apoptotic cells with optic microscope and TEM, treated with 1.25mg/ml OMT for 72h. Exposed 2.5mg/ml OMT for 72h, Many necrosis cells were found. T24 cells treated with OMT, the population of the G0/G1 phase was increased while the population of the S phase T24 cells was declined. 2. The expression of P53, Bcl-2 protein was denclined while the expression of Bax, Caspase-3 protein was increased in T24 cells treaed with OMT, which could induce T24 cells apoptosis.