Effect Of Tyroserleutide On Apoptosis Of Hepatocarcinoma Cell Induced By Mitochondria Injury

Objective: To research the anticancer effect of polypeptides tyroserleutide on humanhepatocarcinoma BEL-7402 and to investigate its possible mechanism by affectingthe function of mitochondria and inducing apoptosis.Methods:1. The inhibiting effect of YSL (100-0.01μg/ml) in vitro on human hepatocarcinomaBEL-7402 and human liver cell L-02 proliferation was assayed by MTT method.2. The effect of YSL on apoptosis of human hepatocarcinoma BEL-7402 and humanliver cell L-02 was assayed by FCM. TEM was used to observe the ultrastructureof human hepatocarcinoma BEL-7402 cell treated with tyroserleutide.3. The-effect of YSL on mitochondrial complexⅣactivity of humanhepatocarcinoma BEL-7402 and human liver cell L-02 was assayed byzymochemistry method.4. The effect of YSL on mitochondrial membrane potential of humanhepatocarcinoma BEL-7402 and human liver cell L-02 was assayed by FCM.5. Western Blot method was used to observe the effect of tyroserleutide onhepatocarcinoma BEL-7402 and human liver cell L-02 active caspase-3expression.Results:1. YSL(0.01-100μg/ml) in vitro can remarkably inhibit human hepatocarcinomaBEL-7402 proliferation. The most effective dosages of YSL are 1μg/ml and10μg/ml. The inhibitory rate of YSL 1μg/ml,10μg/ml dosages are 27.75%,28.47% respectively in 48hrs. YSL was no significantly effect on human liver cell L-02proliferation.2. YSL (1μg/ml,10μg/ml) can significantly induce apoptosis and death of human hepatocarcinoma BEL-7402, the apoptosis rate of YSL 1μg/ml, 10μg/mldosages are 19.95% and 20.18%, the death rate are 47.42% and 40.51%respectively. YSL was no significantly effect on apoptosis of human livercellL-02. Cell ultrastructure shows YSL can induce apoptosis and death of humanhepatocarcinoma BEL-7402 cell, endochylema swelling, edema, mitochondriaand rugh endoplasmic reticulum disaggregates, calcium granulation was seen indillaed endoplasmic reticulum.3. YSL can remarkably decrease the mitochondrial complexⅣactivity of humanhepatocarcinoma BEL-7402, The complexⅣactivity of YSL 1μg/ml,10μg/mldosages are 3.82μmol/min/mg pro and 3.58μmol/min/mg pro respectively in48hrs. YSL was no significantly effect on human liver cell L-02 mitochondrialcomplexⅣactivity.4. YSL can remarkably decrease mitochondrial membrane potentia 1 (ΔΨm) ofhuman hepatocarcinoma BEL-7402 in dosages of 1μg/ml and10μg/ml.ΔΨm didnot change by treated for 0.5hr,ΔΨm began to decrease by treated for 1hr. Thefluorescence intension of all time points are lower than control groups(P＞0.05)during 48hrs. YSL was no significantly effect on mitochondrial membranepotential of human liver cell L-02.5. YSL can promote the protein expression of active caspase-3 of humanhepatocarcinoma BEL-7402. YSL was no effect on the protein expression ofactive caspase-3 of human liver cell L-02.Conclusion: Tyroserleutide can specifically inhibit mitochondrial complexⅣactivityof human hepatocarcinoma BEL-7402, decrease mitochondrial membrane potential,injury tumor cell mitochondria, increase active caspase-3 protein content, inducehuman hepatocarcinoma BEL-7402 apoptosis. YSL was no effect on above-mentioned index of human liver cell L-02.