Kun Zou, Xiaoqin Deng

 Radiotherapy Department, the First Affiliated Hospital of Dalian Medical University, Dalian, China

Objective:The effect of β-elemene on human lungadenocarcinoma A549 cells' hypoxia environment adaptation and radiation inducedEMT. Method: (1). The effect of β-elemene on human lung adenocarcinoma A549 cells proliferation in hypoxia condition. 1. MTT method to detect IC50 ofβ-elemene act on human lung adenocarcinoma cells A549 for 24 h andproliferation inhibition in hypoxia condition. 2. Grouping as follows:control group, hypoxia group, hypoxia with 100µg/ml β-elemene group. After treatment for 48 h, Western Blot deteced the expression of HSP27、NF-κB、HIF-1α、p53protein. (2). The effect of β-elemene on EMT of A549 cell induced by radiation.1. Grouping as follows: control group, 100µg/ml β-elemene group, radiationgroup, radiation with 100µg/ml β-elemene group. Clone formation methods todetect the effect of A549 cell proliferation in each group. 2. Observe the morphological changes of β-elemene on A549 cell EMT formation induced by radiation. 3. Western Blot method to detect the expression of Viminten, N-cadherinand E-cadherin protein in each group. 4. Western Blot method to detect theexpression of HSP27, COX-3, NF-κB, HIF-1α andc-myc protein in each group. 5. Adhesion test to detect the adhesion ability ofA549 cells after different treatments. 6. Wound healing test to detect themigration ability of A549 cells after different treatments. Result: 1. Determined by MTT method to detect that β-elemene has inhibitory effect on the proliferation of A549 cells. Its IC50 is 126µg/ml. With the increase of β-elemene concentration, cell inhibition rate increased gradually under htpoxiacondition. Compared with normoxia group, A549 cells were more sensitive in hypoxia group at low concentrations, but with the increase of β-elemene concentration, the inhibitory effect of A549 cells under hypoxia condition significantly smaller than normoxia group. 2. Hypoxia treatment after 48 h, the expression of HIF-1α、NF-κB、p53、HSP27 protien in A549 cells were increased. After added 100µg/ml β-elemene, the expression of HIF-1α、NF-κB、p53、HSP27protien in A549 cells were decreased compared with separate hypoxia group. 3.The clone number of separate β-elemene group and separate radiation group wereobviously less than control group and the combine group was less than any individual treatment group. 4. After given different radiation dose for 48 h,A549 cellular morphology turned to spindle and extended pseudopodia, intercellularspace also increased. Normal A549 cell was fusiformis and connected closely between each other. A549 cells became EMT change after radiation. With the doseof radiation increased, the number of EMT changed cells increased. After added β-elemene for 24 h, then exposed to radiation another 48 h, the number of EMT changed cells decreased. 5. After radiation, the expression of N-cadherin andViminten were increased and E-cadherin was decreased. After added drug asabove, β-elemene inhibit the change of protein induced by radiation. 6.Compared with the control group, the expression of HSP27, NF-κB, HIF-1α, COX-3and c-myc protein were reduced in radiation group. Moreover, after added drugas above, β-elemene inhibit the change of protein induced by radiation. 7. Withthe radiation dose increased, the adhesion ability of A549 cells alsoincreased. Addedβ-elemene then radiation, the adhesion ability of A549 cells decreased in the same radiation dose. 8. After radiation in the dose of 4 Gy, 8Gy for 48 h, the healing ability of A549 cells increased significantly. Added40µg/ml or 80µg/ml β-elemene then radiation, the healing ability decreased andwas directly proportional to the concentration. Conclusion: 1. β-elemenecan inhibit the proliferation of A549 cells under hypoxia condition by inhibiting the expression of HIF-1α、NF-κB、p53and HSP27 protein. 2. β- elemene may inhibit A549 cell EMT formation byregulating the expression of HSP27, COX-3, NF-κB, HIF-1α, c-myc, Viminten, N-cadherinand E-cadherin protein, thus reduce the migration ability induced by radiation.

KeyWords: A549 cell Hypoxia  EMT β-elemene