The Extraction Of Volatile Oil In Anoectochilus And Its Inhibition Effect In Lung Cancer Cell NCI-H446

Objective: In this research, steam distillation and GC-MS were applied to extract and identify the essential oil from Anoectochilus roxburghii; To study and investigate the essential oil effect hyperplasy inhibit activity toward the the human cell lung cancer cell line NCI-H446.And we further investigated the antitumor mechanism of the steam distillation .Methods: The steam distillation, reflux with water bath extraction and Soxhlet extraction method were applied to investigate the different extraction methods which are index of the essential oil extraction rate; The MTT assay was used to examine the inhibition of NCI-H446 tumor cells induced by the essential oil , and work out its IC50; the essential oil effects on the cell proliferation and apoptosis were observed by transmission electron microscope, TUNEL method and DNA ladder, respectively .Results: The steam distillation of the essential oil extraction rate is 0.06%, which is higher than the other two methods. Compared with other two methods, the steam distillation has the advantage of time-consuming short, less organic solvents volume. GC-MS was used to qualitative analysis of essential oil. The peaks were searched and checked in NIST standard mass spectra library and according to the law of mass spectrometry role, then 37 compounds were identified using the normalization method to calculate the concentration of each compound, accounting for 96.26% of total essential oil. The identified compounds are mainly methyl palmitate and hexadecanoyl acid.The essential oil can remarkably inhibit the proliferation of NCI-H446 tumor cells in a dose-dependent manner, and the IC50 are 70.17,122.85μg/mL after inhibit NCI-H446 tumor cell 24 h and 48 h. After treatment with the essential oil in NCI-H446 tumor cells , the apoptosis increased in a dose-dependent manner , with statistically significant differences from the control group. DNA ladder was seen after the treatment of the essential oil. Conclusion: Anoectochilus roxburghii had high content of essential oil components; the steam distillation is suitable for the essential oil extraction, with short time-consuming, less organic solvents volume and so on. The essential oil can inhibit the proliferation and increase the apoptosis of the human NCI-H446 tumor cells in vitro.