Abstract��OBJECTIVE To study the effect of isoalantolactone on inhibiting proliferation of MCF-7 and induce apoptosis in vitro and further to explore its machinism via mitochondrial and phosphatidylinositol 3-kinase/Akt signaling pathways. METHODS The subject investigated isoalantolactone in MCF-7 cells proliferation inhibition by MTT and SRB methods, and matching the results of two methods; observing morphological changes by inverted microscope and each phase of apoptosis of MCF-7 cells effected by isoalantolactone after 24 h with Hoechst 33258 staining method. Using transmission electron microscopy to observe MCF-7 cells morphology change to determine the mechanism of research; rhodamine 123 tag, laser confocal scanning microscope detection isoalantolactone on the effect of MCF-7 cells mitochondrial membrane potential; Using Western blot to the expression of Bcl-2,Bax,Akt and p-Akt; detecting caspase-3 activity in MCF-7 cells by colorimetry method. RESULTS Isoalantolactone has strong inhibition of proliferation in MCF-7 cells, IC50 values of MTT and SRB methods were 15.21 and 14.908 ��g��mL-1, and in a dose -dependent manner; the morphological of MCF-7 cells was changed after the treatment by Hoechst staining method; morphological changes were observed under transmission electron microscopy of cells display, the drug group cells showed typical apoptotic characteristics of different periods. With the concentrations of isoalantolactone increasing, the level of mitochondrial membrane potential reduced. Western blot result showed that, isoalantolactone could down regulate the expression of anti apoptotic protein Bcl-2, p-Akt, and up regulate the expression of pro-apoptotic protein Bax, had no effect on the expression of Akt protein. And the activity of caspase-3 could be raised by increasing dosage, compared with the control group with significant difference(P<0.01). CONCLUSION Isoalantolactone effectively inhibites the proliferation of MCF-7 cells through mitochondrial and phosphatidylinositol 3-kinase/Akt signaling pathways, which is regulated by activation of caspase-3, down-regulation of Bcl-2, and up-regulation of Bax. Isoalantolactone-induced apoptosis is involved in mitochondrial and the PI3K/Akt pathway.
�����, ����, ������. ����ľ������ͨ���������PI3K/AKTͨ·�յ�MCF-7ϸ���������о�[J]. �й�ҩѧ��־, 2018, 53(8): 601-607.
ZHANG Xiu-juan, LI Lei, HE Li-juan. Effect of Isoalantolactone on Apoptosis in MCF-7 Cells Via Mitochondrial and Phosphatidylinositol 3-Kinase/Akt Signaling Pathways. Chinese Pharmaceutical Journal, 2018, 53(8): 601-607.
LEHMAN C D, ARAO R F, SPRAGUE B L, et al. National performance benchmarks for modern diagnostic digital mammography: update from the breast cancer surveillance consortium[J]. Radiology, 2017,283(1):59-69.
[2]
KOTSOPOULOS J, LUBINSKI J, MOLLER P, et al. Timing of oral contraceptive use and the risk of breast cancer in BRCA1 mutation carriers[J]. Breast Cancer Res Treat, 2014, 143(3): 579-586.
[3]
AGARWAL A, FREEDMAN R A, GOICURIA F, et al. Prior authorization for medications in a breast oncology practice: navigation of a complex process[J]. J Oncol Pract, 2017,13(4):273-282.
[4]
JETHWA K R, KAJILA M M, HUNT K N, et al. Delineation of internal mammary nodal target volumes in breast cancer radiation therapy[J]. Int J Radiat Oncol Biol Phys, 2017, 97(4):762-769.
[5]
TAN R X, TANG H Q, HU J, et al. Lignans and sesquiterpene lactones from Artemisia sieversiana and Inula racemosa[J]. Phytochemistry, 1998, 49(1): 157-161.
[6]
YU H H, LEE J S, LEE K H, et al. Saussurea lappa inhibits the growth, acid production, adhesion, and water-insoluble glucan synthesis of Streptococcus mutans[J]. J Ethnopharmacol, 2007, 111(2): 413-417.
[7]
WEI H, PENG Y, MA G X, et al. Advances in studies on active components of Saussurea lappa and their pharmacological actions[J]. Chin Tradit Herb Drugs(�в�ҩ), 2012, 43(3): 613-620.
[8]
MOGHADAM M H, HAJIMEHDIPOOR H, SAEIDNIA S, et al. Anti-proliferative activity and apoptotic potential of britannin, a sesquiterpene lactone from Inula aucheriana[J]. Nat Prod Commun, 2012, 7(8): 979-980.
[9]
LI Y, LI T K, WEN S W, et al. In vivo and in vitro anti-tumor activity studies of five sequiterpenoids from Inula helenium[J]. Chin Pharmacol Bull(�й�ҩ��ѧͨ��), 2010, 26(1): 112-115.
[10]
KONISHI T, SHIMADA Y, NAGAO T, et al. Antiproliferative sesquiterpene lactones from the roots of Inula helenium[J]. Biol Pharm Bull, 2002, 25(10): 1370-1372.
[11]
KHAN M, DING C, RASUL A, et al. Isoalantolactone induces reactive oxygen species mediated apoptosis in pancreatic carcinoma PANC-1 cells[J]. Int J Biol Sci, 2012, 8(4): 533-547.
[12]
WU M, ZHANG H, HU J, et al. Isoalantolactone inhibits UM-SCC-10A cell growth via cell cycle arrest and apoptosis induction[J]. PLoS One, 2013, 8(9): e76000.
[13]
RASUL A, KHAN M, YU B, et al. Isoalantolactone, a sesquiterpene lactone, induces apoptosis in SGC-7901 cells via mitochondrial and phosphatidylinositol 3-kinase/Akt signaling pathways[J]. Arch Pharm Res, 2013, 36(10): 1262-1269.
[14]
RASUL A, DI J, MILLIMOUNO F M, et al. Reactive oxygen species mediate isoalantolactone-induced apoptosis in human prostate cancer cells[J]. Molecules, 2013, 18(8): 9382-9396.
[15]
KUMAR A, KUMAR S, KUMAR D, et al. UPLC-MS/MS method for quantification and cytotoxic activity of sesquiterpene lactones isolated from Saussurea lappa[J]. J Ethnopharmacol, 2014, 155(2): 1393-1397.
[16]
LEE C H, SHIH Y L, LEE M H, et al. Bufalin induces apoptosis of human osteosarcoma U-2 OS cells through endoplasmic reticulum stress, caspase-and mitochondria-dependent signaling pathyways[J]. Molecules, 2017, 22(3): doi: 10.3390/molecules22030437.
[17]
WANG J, CHENG Y, YIN X, et al. Globular adiponectin inhibits ieptin-stimulated esophageal adenocarcinoma cell proliferation via adiponectin receptor 2-mediated suppression of UHRF-1[J]. Mol Cell Biochem, 2017, 11: doi: 10.1007/s11010-017-2980-6.
[18]
CHANG Q, WANG X L. Cytochrome C, mitochondria and apoptosis[J]. Chin Pharmacol Bull(�й�ҩ��ѧͨ��), 2003, 19(3): 241-244.
[19]
ZHU M Q, WANG C Q. Preparation and in vitro anticance activity of DOX-VES-loaded copolymer micelles[J]. China Pharm J(�й�ҩѧ��־), 2016, 51(23): 2030-2036.
[20]
CHENG J Y, LIU C C, ZENG Z, et al. Shikoninpromotes autophagy of MCF-7 human breast cancer cells through PI3K/Akt pathway[J]. Chin Pharmacol Bull(�й�ҩ��ѧͨ��), 2013, 29(2): 194-198.