Effect Of Chinese Herbal Compound Preparation On Proliferation And Apoptosis Of Skin Cancer A431 Cells

Objective： To investigate the effect of one compound recipe which composed byOphiopogon japonicas、 Pinellia ternate、 Chinese pulsatilla root、 Dandelion、Chuanxiong Rhizoma and Hedyotis diffusae on the proliferation and apoptosisinduction in epidermal squamous cell carcinoma cell line A431in vitro.Methods: Humane Pidermal squamous cell carcinoma cell line A431wascultured.After the treatment with the compound recipe,inhibition of cellular growthwas determined by measuring MTT dye absorption of the living cells. Lightmicroscopy observed morphological changes. And lactate dehydrogenase (LDH) in thecell were measured by spectroscopy respectively. The compound recipe induced A431cell apoptosis was assessed with Hochest33258nucleus fluorescent staining. Cytc wasdetected by elisa and both of Bax and Bcl-2were detected by immunohistochemistry oncells. The DNA content and apoptosis were assessed by flowey tometry.Results：MTT assay was applied to survey proliferation-inhibitory effect of baicaleinon A431cell, normal-optic-microscope was employed to observe cell growth and theirMorphologic change.Light microscopy revealed characteristic morphological changesof cell apoptosis. The level of LDH,Caspase-3，Caspase-9and Cytc were increasedsignificantly compared with the negative control group. With Hochest33258staining,the nucleus in the compound recipe group showed a dark blue fluorescence,with the appearance of typical morphological features of apoptosis, such askaryopyknosis and apoptotic body, etc. The expression of Bax of A431cells wasincreased and the expression of Bcl-2was decreased after treatment of the compoundrecipe,which was statistically different from the negative control group. The curcumingroups can obviously prompt the apoptosis rate, and compared with the negativecontrol group, the cells in G0-G1phase were higher,and cells in the G2-M phase werelower.Conclusion： The compound recipe plays an inhibitory role in the tumor cellproliferation. The active mechanisms may be include:(1) Destory cell membrance, andincrease the release of cytochrome C、Caspase-3and Caspase-9.(2) Enhance the expression of Bax，reduce the expression of Bcl-2to interfere with mitochondrialapoptosis signal transduction pathway to induce apoptosis.