Five Kinds Of Traditional Chinese Medicine Anti-NPC In Vitro And SDH Extract Induced Apoptosis Of NPC Cells

Background and Objective: Nasopharyngeal carcinoma (NPC) is com-mon in southeast of China, and the 5-year-survival rate is only 50%～70% after radiotherapy and chemotherapy, radioresistance, chemoresistance and severe complications lower the effect of combining treatment for nasopharyngeal car-cinoma. The search for more effective and less toxic side effects of anti-NPC drugs and improve efficacy of clinical treatment is an urgent need to address the problem. Traditional Chinese medicine (TCM) is that people constantly sum up, selection derived at life and disease, proven to be effective. We study the sensi-tivity of human NPC cells to five kinds of common TCM in Guangxi through NPC Chemosensitivity Assay in vitro, and anti-NPC mechanisms of the best one through Cell biology and molecular biology.Method: Extraction of five kinds of TCM ingredients respectively, includ-ing Scutellaria barbata D.Don(SB), Hedyotis diffusa(HD), Herba Agrimo-niae(HA), Nitidine(ND), Selaginella doederleinii Hieron(SDH). CNE1 and CNE2 NPC cell were treated with these five Liquid Extracts and drug containing serum at different concentration, serum pharmacology and Methyl Thiazolyl Tetrazolium (MTT) assay in vitro were used to measure the inhibition rate of drugs on growth of NPC cells. TW03, HONE1 NPC cells and HFL-I human embryonic lung fibroblast cell were treated with SDH at different concentration, MTT assay was used to observe the effect of SDH on growth of three cell lines; Phase contrast microscope was used to observe the cells morphology changed. The change on cell cycle was analyzed by Propidium Iodide (PI) staining and flow cytometry (FCM). Annexin V-FITC and PI staining FCM method was used to observe the apoptosis of the NPC cells. The expression changes of Bax and Bcl-2 in NPC cells induced by SDH were detected by Immunohistochemistry.Results: SDH Extract and SDH containing serum significantly inhibited the growth and Proliferation of CNE1, CNE2 cells in time and dose dependent manner. A certain concentration of ND Extract and ND containing serum could inhibit the growth of CNE1, CNE2 cells. High concentration of SB, HD and HA Extracts could inhibit the growth of CNE1, CNE2 cells, while their containing serums couldn't. 2.5mg/mL SDH Extract could inhibit the growth of human NPC cell obviously in vitro, while could inhibit the growth of HFL-I cell little. NPC cell growth was inhibited and cell death increased was observed under phase contrast microscope. SDH Extract affected NPC cell cycle distribution and arrested cell in S phase; in Annexin V-FITC and PI staining FCM method, SDH Extract could promote NPC cell apoptosis, and this effect of SDH Extract was in a dose-dependent manner. in NPC cell the expression of Bax was higher and Bcl-2 was lower when NPC cell was treated with SDH Extract 24h meas-ured by immunohistochemistry.Conclusions: SDH Extract and containing serum can inhibit the growth of human NPC cell obviously in vitro. Selaginella doederleinii Hieron Extract and containing serum Probably belong to cell cycles specific agents. Selaginella doederleinii Hieron extract can induce apoptosis on NPC cell efficiently. Bax and Bcl-2 are very important apoptosis regulation protein , and play a role in the apoptosis of NPC cells treated by Selaginella doederleinii Hieron extract.