protein levels of PI3K/Akt/ HIF-1 pathway and key enzymes involved in glycolysis

 
X.Zhang,etal. Cancer Letters 481 (2020) 32–44 Fig. 5. SSL6 suppresses glycolysisvia blocking CD47/PI3K/Akt/HIF-1 signaling pathway. (A) After 72 h of SSL6 treatment (10 μg/mL), protein levels of PI3K/Akt/ HIF-1 pathway and key enzymes involved in glycolysis in Huh-7 were determined. (B) After 72 h of SSL6 treatment (10 μg/mL), protein levels of PLCγ and phosphorylated-PLCγ were assessed. (C) Huh-7 cells were treated with SFN (5 μM) and/or SSL6 (10 μg/mL) for 72 h, the expression of PKA, AMPK and phos- phorylated-AMPK were examined. 

(D,E) After knockdown of CD47, protein levels of PI3K/Akt/HIF-1 pathway and key enzymes in glycolysis in Huh-7 (D) and MHCC97H (E) cells. (F) Flow cytometry detection of cell death after Huh-7 cells were pre-treated with LY294002 (20 μM) or 2-DG (2.5 mM) for 12 h, SFN (5 μM) and/orSSL6(10μg/mL)wereadministratedfor72h.(G,H)Huh-7cellswerepre-treatedwithLY294002(20μM),theintracellularlactatelevels(G)andthemRNA expression of CD47,HIF1A and key enzymes involved in glycolysis (H) were determined. Data are expressed as means ± SEM (n = 3). *p < 0.05, **p < 0.01, ***p < 0.001.NS denotes no significance. for SFN-induced CD47 but SFN might also inhibit CD47 via a PI3K- 3.6. SSL6reversesthesensitivityofSFN-resistant(SFN-R)HCCcells independent pathway when PI3K was blocked. Together these results illustrate that SSL6 down-regulates glycolysis and sensitizes HCC cells We next wondered whether SSL6 could re-sensitize SFN-R HCC to SFNvia CD47/PI3K/Akt/HIF-1 signaling pathway. cells. We established SFN-R Huh-7 and MHCC97H cells by continuous induction with low concentration of SFN for more than 1 month (The 39
X.Zhang,etal. Cancer Letters 481 (2020) 32–44 Fig. 6. SSL6 re-sensitizes SFN-resistant (SFN-R) HCC cells to SFN.

 (A) Schematic models of SFN-R HCC cell induction. (B) IC50 of control and SFN-R Huh-7 and MHCC97H cells. (C)CD47 mRNA expression in control and SFN-R HCC cells (Huh-7 and MHCC97H). (D) Flow cytometry detection of CD47 expression on control and SFN-R HCC (Huh-7 and MHCC97H). (E) Western-blot of CD47 with control and SFN-R MHCC97H cells. (F) Lactate levels in control and SFN-R HCC cells (Huh-7 and MHCC97H). (G) SFN-R Huh-7 cells were treated with SFN (7.5 μM) and/or SSL6 (10 μg/mL) for 72 h, the cell death were de###http://www.glpbio.com/simage/GA11233-L-NAME-hydrochloride-4.png####termined with flow cytometry. Data are expressed as means ± SEM (n = 3). *p < 0.05, **p < 0.01, ***p < 0.001. IC50 values for SFN in Huh-7 and Huh-7 SFN-R cells were 4.720 and (Fig. 6C Huh-7 cells: the increase percentage of SFN-Rvs. control was 7.060 μM, respectively; in MHCC97H and MHCC97H SFN-R cells were 62 Cell Counting Kit-8 glpbio.64%,p=0.0092;MHCC97Hcells:theincreasepercentageofSFN-R 8.31 and 16.28 μM, respectively) (Fig. 6A and B). These SFN-R cells vs. control was 34.39%, p = 0.0006. Fig. 6D Huh-7 cells: the increase showed increased CD47 expression and elevated glycolysis levels percentage of SFN-R vs. control was 88.99%, p < 0.0001; MHCC97H 40
X.Zhang,etal. Cancer Letters 481 (2020) 32–44 Fig. 7. SSL6 and SFN synergistically attenuate xenograft HCC growth in vivo. 

(A) Protocol of in vivo experiment model BCI-121. (B) Curves of tumor growth in groups of control (PBS), SSL6 (10 mg/kg), SFN (2 mg/kg), and SFN (2 mg/kg) plus SSL6 (10 mg/kg). (C) Tumor entity view from groups of control (PBS), SSL6 (10 mg/kg), SFN(2mg/kg),andSFN(2mg/kg)plusSSL6(10mg/kg).(D)IHCforCD47expressionintumorscontrol(PBS),SSL6(10mg/kg),SFN(2mg/kg),andSFN(2mg/kg) plus SSL6 (10 mg/kg) 3x flag peptide. cells: the increase percentage of SFN-R vs. control was 35.04%, percentage of SFN vs. control was 177.85%, p = 0.0018; the increase p = 0.0008) (Fig. 6C–F and Supplementary Fig. 4A). In addition, SSL6 percentage of SSL6vs. control was 108.27%, p = 0.0191; the increase promoted the death of SFN-R Huh-7 and MHCC97H cells, as well as percentage of SFN + SSL6vs. SFN was 55.65%, p = 0.0054) (Fig. 6G enhanced SFN-induced cell death (Fig. 6G Huh-7 SFN-R: the increase and Supplementary Fig. 4B). 41

Comments

Post a Comment

Popular posts from this blog

Zhao, et al. Cancer Letters 481 (2020) 15–23 Y. Ohsumi, T. Tokuhisa, N. Mizushima

 C. Zhao, et al. Cancer Letters 481 (2020) 15–23 Y. Ohsumi, T. Tokuhisa, N. Mizushima, The role of autophagy during the early of autophagosome-lysosome fusion in human cancer cells, Autophagy 8 (2012) neonatal starvation period, Nature 432 (2004) 1032–1036. 338–349 Diphenylterazine (DTZ) . [11] S.H. Noh, S.R. Park, H.K. Yang, H.C. Chung, I.J. Chung, S.W. Kim, H.H. Kim, [30] J. Li, N. Hou, A. Faried, S. Tsutsumi, T. Takeuchi, H. Kuwano, Inhibition of au- J.H. Choi, H.K. Kim, W. Yu, J.I. Lee, D.B. Shin, J. Ji, J.S. Chen, Y. Lim, S. Ha, tophagy by 3-MA enhances the effect of 5-FU-induced apoptosis in colon cancer Y.J. Bang, C.t. investigators, Adjuvant capecitabine plus oxaliplatin for gastric cells, Ann. Surg Oncol. 16 (2009) 761–771. cancer after D2 gastrectomy (CLASSIC): 5-year follow-up of an open-label, rando- [31] K.N. Dalby, I. Tekedereli, G. Lopez-Berestein, B. Ozpolat, Targeting the prodeath mised phase 3 trial, Lancet Oncol. 15 (2014) 1389–1396. and prosurvival f...
Read more

Zhao, et al. Cancer Letters 481 (2020) 15–23 (caption on next page) 20

 r> C. Zhao, et al. Cancer Letters 481 (2020) 15–23 Fig. 5. The combination of Prodigiosin and 5-Fu inhibits the growth of HCT116 xenografts in nude mice. Nude mice bearing HCT116 xenograft tumors were treated with vehicle, prodigiosin alone (1 mg/kg), 5-Fu alone (30 mg/ kg), or a combination of prodigiosin and 5-Fu twice weekly through i.p. injection for 21 days after inocula- tion with cultured tumor cells. (A) Tumor images and tumor weights are shown.Onday21afterinoculation,the mice were sacrificed, and the tumor tissues were imaged, weighed, and de- tailed. Mean ± SD (n = 5); **, P < 0.01 versus control, prodigiosin or 5-Fu alone. (B) Tumor volume was recorded every three days after treat- ment. Mean ± SD (n = 5); **, P < 0.01 versus control, prodigiosin or 5-Fu alone.  (C) Body weight was recorded every three days during the 21 day of exposure. No sig- nificant differences were detected be- tween the vehicle and other groups: prodigiosin alone CORM-3 , 5...
Read more

Prodigiosin Inhibition of autophagy is a promising strategy to augment the cytotoxicity of chemotherapeutic agents

 Cancer Letters 481 (2020) 15–23 Contents lists available at ScienceDirect Cancer Letters journal homepage: www.elsevier.com/locate/canlet Prodigiosin impairs autophagosome-lysosome fusion that sensitizes T colorectal cancer to 5-fluorouracil-induced cell death a,b,1 a,b,1 a,b a,b a,b a,b Chong Zhao , ShaoZhuang Qiu , Jie He , Yao Peng , Haoming Xu , Zhiqiang Feng , a,b a,b a,b a,b,∗ Hongli Huang , Yanlei Du , Yongjian Zhou , Yuqiang Nie a Department of Gastroenterology, Guangzhou Digestive Disease Center, Guangzhou First People\'s Hospital, Guangzhou Medical University, Guangzhou, 510180, China b Department of Gastroenterology, Guangzhou First People\'s Hospital, School of Medical, South China University of Technology, Guangzhou, 510180, China ARTICLE INFO ABSTRACT Keywords: Chemotherapy failure is a major cause of recurrence and poor prognosis in colorectal cancer (CRC) patients. Prodigiosin Inhibition of autophagy is a promising strategy to augment the cytotoxicit...
Read more