Colorectal Surgery
Influences of autophagic regulations of carcinoma associated fibroblasts on the migration and invasion of cocultured colorectal cancer
Wei Peng, Changsong Ma, Fuhua Han, Chungen Xing, Yongyou Wu
Published 2018-02-08
Cite as Chin J Exp Surg, 2018, 35(2): 230-233. DOI: 10.3760/cma.j.issn.1001-9030.2018.02.011
Abstract
ObjectiveTo explore the influences of autophagic regulations of carcinoma associated fibroblasts (CAFs) on the migration and invasion of cocultured colorectal cancer.
MethodsCAFs were attempted by primary cultured tissues from colorectal cancer. Autophagic regulation of CAFs was performed using rapamycin and bafilomycin, and autophagic effects were observed with anti-microtubule-associated protein 1 light 3 (LC3). The influences of autophagic regulation of CAFs on the migration and invasion abilities of CCL244 were observed by transwell invasion assay and scratch assay. Western blotting analysis of tumor protein 53 (p53), B-cell lymphoma/leukemia-2 (bcl-2), bcl-2 assaciated X protein (bax), nuclear factor-kappa B (NF-κB) and cysteine-containing aspartate-specific proteases-3 (Caspase-3) were performed determined to investigate the potential mechanisms.
Results400 nmol/L rapamycin increased autophagy and the expression of autophagy associated proteins damage-regulated autophagy modulator (DRAM), autophagy associated gene 5 (Atg5), Beclin-1 and LC3 was up-regulated.400 nmol/L bafilomycin inhibited autophagy of CAFs and the above proteins were down-regulated. We find that rapamycin pretreatment CAFs+ CCL244 group has most tumor cells to across the well (172.0±9.3) and bafilomycin pretreatment CAFs+ CCL244 group has least tumor cells to across the well (52.0±5.0) after each group cells were cultured in transwell in 48 h. The migration rate of CCL-244 cells pretreated in Rapamycin [(15.34±0.49) mm] was especially higher than the bafilomycin pretreatment group, There was no significant difference between the control group and bafilomycin pretreatment group [(10.32±1.08) mm]. Induction of autophagy in CAFs upregulated the expression of bcl-2 and NF-κB and downregulated the expression of bax, p53 and Caspase-3 in irradiated CCL244. Inhibition of autophagy in CAFs showed the opposite results in CCL244.
ConclusionUp regulation of autophagy in CAFs can enhance the ability of invasion and migration of cocultured cancer cells, while inhibition of autophagy in CAFs can reduce the invasion and migration ability of cocultured cancer cells.
Key words:
Colorectal cancer; Carcinoma associated fibroblasts; Autophagy; Regulation; Migration
Contributor Information
Wei Peng
Department of General Surgery, the Second Affiliated Hospital of Soochow University, Suzhou 215004, China
Changsong Ma
Fuhua Han
Chungen Xing
Yongyou Wu
