RESUMO
The main objective of the present research work was to evaluate the antitumor effects of ethanol extract of Semen Euphorbiae (EESE) in ACHN human renal carcinoma cells. The effects on apoptosis induction, cell cycle phase distribution and livin protein expression were also evaluated. MTT assay was used to assess cytotoxic effects of the extract while as clonogenic assay was used to evaluate effects on colony formation tendency. Inverted phase contrast and fluorescence microscopic techniques were used to evaluate effects of EESE on cellular morphology and apoptosis. Flow cytometry using annexin V-FITC and propidium iodide were used to quantify the extent of apoptosis and also to evaluate effects on cell cycle. Results indicate that ethanol extract of Semen Euphorbiae exhibited potent cytotoxic effects in ACHN human renal cancer cells. These effects were found to be dose-dependent as well as time dependent. Clonogenic assay revealed that EESE led to dose-dependent inhibition of colony formation in these cells. EESE-treated cells also showed evident signs of alterations and deformations in cell morphology including detachment of cells from one another forming small cluster of cells. In contrast to untreated control cells, EESE-treated cells with 10, 100 and 200 µg/ml dose showed an increase in the number of cells emitting red/orange fluorescence indicating onset and execution of apoptosis. EESE extract also led to G2/M cell cycle arrest in these cells.
RESUMO
BACKGROUND Cervical cancer is one of the major causes of cancer death of females worldwide. Radiotherapy is considered effective for cervical cancer treatment, but the low radiosensitivity found in some cases severely affects therapeutic outcomes. This study aimed to reveal the role of CD146, an important adhesion molecule facilitating tumor angiogenesis, in regulating radiosensitivity of cervical cancer cells. MATERIAL AND METHODS CD146 protein expression was compared in normal cells, cervical cancer cells with lower radiosensitivity, and cervical cancer cells with higher sensitivity from cervical squamous cell carcinoma patients. Anti-CD146 monoclonal antibody AA98 was used to inhibit CD146 in human cervical cancer SiHa cells with relatively low radiosensitivity, and then the cell survival and apoptosis changes after radiation were detected by colony formation assay and flow cytometry. RESULTS CD146 protein was significantly up-regulated in cervical cancer cells (P<0.001), especially in cancer cells with lower radiosensitivity. The SiHa cells treated with AA98 showed more obvious inhibition in cell survival (P<0.05) and promotion in cell apoptosis (P<0.01) after radiation, compared to the untreated cells. More dramatic changes in apoptotic factors Caspase 3 and Bcl-XL were also detected in AA98-treated cells. CONCLUSIONS These results indicate that inhibiting CD146 improves the effect of radiation in suppressing SiHa cells. This study shows the potential of CD146 as a target for increasing radiosensitivity of cervical cancer cells, which might allow improvement in treatment outcome in cervical cancer. Further studies are necessary for understanding the detailed mechanism of CD146 in regulating radiosensitivity.