Pancratistatin Inhibits the Growth of Colorectal Cancer Cells by Inducing Apoptosis, Autophagy, and G2/M Cell Cycle Arrest
Authors: Dong-Yang Liu
Yi-Jia Xiong
Rong-Rong Shen
Yong Xiong
Journal: Medical Science Monitor (2019)
Discipline: CLINICAL MEDICINE
Keywords: colorectal cancer; pancratistatin; apoptosis; cell lines; cell viability
Affiliations: Department of General Surgery, Shanghai Sixth Peoples' Hospital East Affiliated to Shanghai University of Medicine and Health Sciences, Shanghai, China (mainland)., Department of Radiology, Shanghai Sixth Peoples' Hospital, Shanghai, China (mainland).

Abstract:  BACKGROUND Worldwide, colorectal cancer is ranked as the third most prevalent cancer. The natural compound, pancratistatin, extracted from the spider lily, has previously been shown to target apoptosis in cancer cells lines. This study aimed to investigate the effects of pancratistatin in human colorectal cancer cells in vitro. MATERIAL AND METHODS Human colorectal cancer cell lines, including HTC-15 cells, were compared with a normal human colonic fibroblast cell line, CDD-18Co. Cells were treated with increasing doses of pancratistatin. The MTT assay was used to assess cell viability. Fluorescence microscopy using DAPI and Annexin-V/propidium iodide (PI) was used to detect cell apoptosis. Cell autophagy was detected by electron microscopy. Cell migration was evaluated using a wound healing assay, and Western blot determined the expression levels of cell cycle proteins. RESULTS Pancratistatin inhibited the growth of the colorectal cancer cells with an IC₅₀ ranging from 15-25 µM, but had a limited effect in normal CCD-18Co cells, with an IC₅₀ of &gt;100 µM. Pancratistatin reduced HCT-15 cell migration. Growth inhibition due to pancratistatin was associated with morphological changes of HCT-15 cells and included autophagy and apoptosis, and increased expression the autophagic proteins, LC3II, beclin-1, and Bax. Pancratistatin induced arrest of HCT-15 cells at G2/M of the cell cycle and inhibited phosphorylation of cdc2/cyclin-dependent kinase 1 (CDK1) and Cdc25c and the expression of cyclin B1. CONCLUSIONS Pancratistatin inhibited the growth of colorectal cancer cells in vitro by inducing apoptosis, autophagy, and G2/M cell cycle arrest. 

Retracted: Yes

PubPeer comments:
 *[right]* Fig from [Xiong et al (2019)] .  Western blot shows that pancratistatin resulted in increased expression of Bax and reduced expression of Bcl-2. *[left]* Fig 6C from [Zhang et al (2018)] .  Effect of ludartin at the IC50 of the Saso-2 osteosarcoma cells. .... (C) Expression of MMP-2 and 9 by Saso-2 osteosarcoma cells . ![file] , 
 #1 *[right]* Fig from [Xiong et al (2019)] .  Western blot shows that pancratistatin resulted in increased expression of Bax and reduced expression of Bcl-2. *[left]* Figure 6B from [Zhang, Zhang Ni (2020)] .  Effect of indicated doses eriodictyol on ... (B) expression of Bax Bcl-2 by western blotting . *[centre]* Fig 6C from [Zhang et al (2018)] .  Effect of ludartin at the IC50 of the Saso-2 osteosarcoma cells. .... (C) Expression of MMP-2 and 9 by Saso-2 osteosarcoma cells . ![file] UPDATE: Earliest sighting so far is Figure 5 from [Huang et al (2017)] .  Effect of EGCG at indicated concentrations showing the expression of Bax and Bcl-2 in A549 cells as determined by Western blotting. ... The results showed that EGCG caused upregulation of Bax and downregulation of Bcl-2 in a concentration-dependent manner. ![file] , 
 *[left]* Figure 3 from [Xiong et al (2019)] .  Annexin-V propidium iodide (PI) staining shows the percentage of the apoptotic HCT-15 cells at indicated  . *[right]* Figure 2B from [An Liu (2020)] .  Camelliol C induces apoptosis in cervical cancer cells. ... annexin V PI (B) staining of camelliol C-treated HeLa cells, showing induction of apoptosis . ![file] , 
 *[left]* Figure 4 from [Xiong et al (2019)] .  Western blot shows that pancratistatin treatment resulted in a dose-dependent increase in the expression of LC3 II, ATG-12, ATG-5, and beclin-1. *[right]* Fig 4b from [Li et al (2020)] .  Efect of Indirubin on the expression of autophagy related proteins. ![file] , 
 Figure 7B.  The effects of pancratistatin on the cell cycle and expression of cell cycle-associated proteins in colorectal cancer cells in vitro. .. (B) Western blot shows that pancratistatin is associated with a dose-dependent decrease in the expression of the cell cycle-associated proteins . ![file] Colored rectangles are mine, to facilitate comparison with *[left]* Figure 8 of [Pan Zhang (2019)] .  Effect of indicated concentrations of Linalool on the PI3K AKT signalling pathway as indicated by western blotting. The Figure depicts that Linalool blocks the PI3K AKT pathway in OECM-1 cells concentration-dependently. *[right]* Figure 5 from [Zhang et al (2018)] .  Western blots showing the effect of ludartin on cell cycle proteins in Saso-2 osteosarcoma cells. (A) Cdc25c, pCdc25c, Cdc2, pCdc2, p21WAFI, and cyclin B1, and (B) p21, p53, and p27. Ludartin treatment increased the expression of cyclin-dependent kinase inhibitor p21 (WAF1) or p21WAF1 in Saso-2 osteosarcoma cells but did not alter the expression of p27 or p53 tumor suppressor proteins . ![] Familiar from Fig 8A,B of [Luo et al (2019)] : ![] , 
 Retracted [25 March 2021] .  An editorial decision has been made to retract this manuscript due to breach of publishing guidelines, following the identification of non-original and manipulated figures. , 
 #2 ![] The ultimate source of this versatile loading band appears to be Fig 5A from  Schisandrin B induces apoptosis and cell cycle arrest of gallbladder cancer cells ([Xiang et al 2014] ).  NOZ cells were treated with Sch B (0, 30, 60, and 90 u03bcmol L) for 48 h. The expression levels of Bcl-2, Bax, cleaved caspase-9, cleaved caspase-3, cleaved PARP and NF-u03baB were detected by western blot analysis . ![file] , 
 #4 *[left]* Figure 4 from [Xiong et al (2019)] .  Western blot shows that pancratistatin treatment resulted in a dose-dependent increase in the expression of LC3 II, ATG-12, ATG-5, and beclin-1. *[right]* Fig 4b from [Li et al (2020)] .  Effect of Indirubin on the expression of autophagy related proteins. ![file] The source (or the earliest sighting) is Fig 3b from  Escin induces caspase-dependent apoptosis and autophagy through the ROS p38 MAPK signalling pathway in human osteosarcoma cells in vitro and in vivo ([Zhu et al 2017] ). ![file] ,