Alantolactone inhibits cell autophagy and promotes apoptosis via AP2M1 in acute lymphoblastic leukemia

Abstract

Background Acute lymphoblastic leukemia (ALL) is an aggressive hematopoietic malignancy that is most commonly observed in children. Alantolactone (ALT) has been reported to exhibit anti-tumor activity in different types of cancer. The aim of the present study was to investigate the anti-tumor activity and molecular mechanism of ALT in ALL. Methods ALL cell lines were treated with 1, 5 and 10 μM ALT, and cell viability was assessed using an MTT assay and RNA sequencing. Flow cytometry, JC-1 staining and immunofluorescence staining assays were used to measure cell apoptosis and autophagy. Additionally, western blot analysis was used to detect expression of apoptosis and autophagy related proteins. Finally, the effects of ALT on tumor growth were assessed in a BV173 xenograft nude mouse model. Results ALT inhibited the proliferation of ALL cells in a dose-dependent manner. Additionally, it was demonstrated that ALT inhibited cell proliferation, colony formation, autophagy, induced apoptosis and reduced tumor growth in vivo through upregulating the expression of adaptor related protein complex 2 subunit mu 1 (AP2M1). Moreover, the autophagy activator rapamycin, attenuated the pro-apoptotic effects of ALT on BV173 and NALM6 cell lines. Overexpression of AP2M1 decreased the expression of Beclin1 and the LC3-II/LC3-1 ratio, and increased p62 expression. Knockdown of Beclin1 increased the levels of bax, cleaved caspase 3 and cytochrome C, and decreased bcl-2 expression. Conclusions The present study demonstrated that ALT exerts anti-tumor activity through inducing apoptosis and inhibiting autophagy by upregulating AP2M1 in ALL, highlighting a potential therapeutic strategy for treatment of ALL.

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“…Apart from IAL [ 202 ], recently particularly ALT has been extensively studied for its cytotoxic and anti-proliferative activity in a variety of human cancer cell lines, including the colon [ 203 ], lung [ 204 ], breast [ 205 ], gastric [ 206 ], pancreatic cancer [ 207 ], leukemia [ 208 ], and glioblastoma multiforme [ 209 ]. The anti-tumor properties of ALT occurred through a wide range of overlapping molecular pathways, interacting with each other [ 180 , 207 ].…”

“…The inhibitive effect of ALT treatment (20 μM) on TrxR1 activity and induction of ROS-mediated p38 MAPK signaling pathway in gastric cancer cells could recently be shown [ 206 ]. Moreover, Shi and colleagues were the first to report induced apoptosis by ALT (1.5 and 10 μM) in a dose-dependent manner and the inhibition of autophagy by upregulating AP2M1 in acute lymphoblastic leukemia [ 208 ]. Only recently, it was found that ALT inhibited the metastatic phenotype and induced the apoptosis in glioblastoma cells by targeting LIMK kinase activity and activating the cofilin/G-actin signaling cascade [ 210 ].…”

“…Moreover, an anti-cancer effect of ALT treatment on esophageal cancer cells through inhibition of the Wnt/β-catenin signaling pathway was reported [ 211 ]. ALT also exerted synergistic anti-tumor effects with other chemotherapeutics [ 208 ]. For instance, ALT strengthened the efficacy of epidermal growth factor receptor inhibitors such as erlotinib or afatinib against pancreatic cancer cells [ 207 ], as well as cisplatin against lung adenocarcinoma [ 204 ].…”

Alantolactone: A sesquiterpene lactone with diverse pharmacological effects

“…In the past two decades, the most significant progress in the researches of Ala has been its anticancer activity. Since 1976 when Ala first showed cytotoxicity against isolated human leukocytes (Dupuis & Brisson, 1976), the apoptosis-inducing effects in leukemia cells have been reported by many groups (Ahmad et al, 2021;Ding, Gao, et al, 2016;Dirsch et al, 2001 2019; Yang et al, 2013;, with models including Jurkat leukemia T cells (Dirsch et al, 2001;Xu et al, 2019), HL-60 cells (Ding, Gao, et al, 2016;Pal et al, 2010;Xu et al, 2019), RPMI-8226 cells , K562 cells (Ding, Gao, et al, 2016;Wei et al, 2013;Xu et al, 2019;Yang et al, 2013), multiple myeloma cells , KG1a cells (Ding, Gao, et al, 2016), BV173 cells (Shi et al, 2020), NALM6 cells (Shi et al, 2020), JM-1 cells (Shi et al, 2020), NALM1 cells (Shi et al, 2020), RS6 cells (Shi et al, 2020), SUPB15 cells (Shi et al, 2020), and THP-1 cells (Ahmad et al, 2021;Ding, Gao, et al, 2016), and the anticancer spectrum has also been broaden to liver cancer (Kang et al, 2019;Khan et al, 2013;Lei et al, 2012;Mi et al, 2014;Zhang, Xu, et al, 2016), glioblastoma (Khan et al, 2012;Wang et al, 2017), colon/colorectal cancer (Ding, Wang, et al, 2016;Shi et al, 2011;Zhang, Xu, et al, 2016;Zhang et al, 2013), breast cancer (Chun et al, 2015;…”

“…Ala treatment can increase the expression of adaptorrelated protein complex 2 subunit μ1 (AP2M1), which results in the higher level of p62 (autophagy substrate), and the lower levels of Beclin 1 and LC3-II/LC3-I ratio (autophagy markers) (Shi et al, 2020). The attenuated autophagic degradation may result from the reduced expression of transcription factor EB (TFEB) and lysosomal alkalinization, which indicate the dysfunction of lysosomes, because TFEB and its associated hydrolases cathepsin B and cathepsin D regulates lysosomal biogenesis and function (He et al, 2018).…”