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[Effect of cigarette smoke extract on RAW264. 7 cell proliferation, autophagy and its mechanism]
Manli Wang1, Jincheng Liu1, Min Mu1
1Frontier Experimental Center of Medical College, Anhui University of Science and Technology;Department of Preventive Medicine, Medical College, Anhui University of Science and Technology, Huainan 232000, China.
Objective:
To inquiry the effects of cigarette smoke extract(CSE) on RAW264. 7 cell proliferation, autophagy and its mechanism.
Methods:
RAW264. 7 cell were used and divided into control, starvation and CSE group(2%, 3%, 4%, 5%CSE). CCK-8 was used to detect the toxic action of CSE on RAW264. 7 cell. Western blot and mRFP-GFP-LC3 cell fluorescence spot count were used to explore the function of CSE on RAW264. 7 cell autophagy and its mechanism.
Results:
Compared with the control group, the result of CCK-8(0. 671 ± 0. 03、0. 746± 0. 10、0. 584 ± 0. 07、0. 588±0. 05) showed that CSE inhibit the proliferation of RAW 264. 7 cell on 24 hours, the difference was statistically significant(P < 0. 05). The outcomes of Western blot showed that, compared with the control group, LC3 B in the CSE group increased, difference in 6(6. 612 ± 0. 35)/12(4. 383 ± 1. 99)/24(5. 781 ± 0. 78) hours, while P62 decreased in 6(1. 815±0. 08)/12(4. 383±1. 99)/24(0. 414±0. 06) hours also different, P-mTOR(1. 744 ± 0. 15) and P-AKT(0. 376 ± 0. 03) decreased, the difference was statistically significant(P<0. 05), but Beclin1 was not significantly changed. The mRFP-GFP-LC3 cell fluorescence spot count showed that the green fluorescence spot(GFP)decreased and the red fluorescence spot(mRFP) remained stable in CSE group, combined mRFP-GFP is shown as yellow and red spots.
Conclusion:
CSE has toxic effect on cell proliferation and leads to RAW264. 7 cell autophagy enhanced through AKT/m TOR pathways.
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