Objective: To study the effect of apoptosis induction of pianzihuang in human osteosarcoma U-20S cells.
Methods: Sixth SD rats were randomly divided into group A (8 rats) and group B (8 rats) in the study. Among them, 8 rats (group A) were treated with PBS and 8 rats (group B) treated with pianzihuang. (2) Human osteosarcoma U-2OS cells were cultured in nutrient solution (including fetal calf serum of 10%, penicillin of 100 IU/ml, streptomycin of 100 microg/ml), the environmental condition in saturated humidity 5% CO2, 37 degrees C. The osteosarcoma U-2OS cell proliferation effect by different saturation of blood serum was measured by MTT method. And the optimal concentration of the serum (including Pianzihuang) could be obtained. (3) The morphous of osteosarcoma U-2OS cell were observed by inverted phase contrast microscope and electron microscope. (4) Apoptosis cell was detected by TUNEL in situ end labeling. (5) Extracted DNA,agarose gel electrophoresis (AGE) to detect apoptosis cell by transmission electron microscope (TEM).
Results: (1) It was best that osteosarcoma U-2OS cell proliferation effect by 20% of the concentration of the serum (including Pianzihuang). (2) On inverted phase contrast microscope showed that the cells of group A were fusiform adherence growth; the cyton transform shrind and circle in 20% of the concentration of the serum (including Pianzihuang). Follow lasting time,the cell turning circle gradually increased. Necrosis cell and fragment were found by microscope. (3) The detected result of apoptosis cell in 20% of the concentration of the serum(including Pianzihuang) was positive by TUNEL assays. (4) AGE showed "ladder" strand of DNA, a special phenomenon of cellular apoptosis. (5) Chromatin aggregated along corrugativus caryotheca and apoptotic body appeared on cell surface by observation of TEM.
Conclusion: Pianzihuang can inhibit proliferation and induce apoptosis for osteosarcoma U-2OS cell.