DNAJB6b通过激活AKT通路增强结直肠癌细胞的侵袭迁移能力

doi:10.3969/j.issn.1004-616x.2022.03.003

Abstract

Abstract: OBJECTIVE: To investigate effects from over-expression of DnaJ heat shock protein family member B6 isoform b (DNAJB6b) on invasive and migration abilities of colorectal cancer (CRC) cells. METHODS: Expressions of DNAJB6b in cultured CRC cell lines DLD-1 and HCT116 were knocked down with small interfering RNA (siRNA) and short hairpin RNA (shRNA),respectively.Control cells were transfected with negative control siRNA and shRNA.Changes in protein levels were detected by Western blot.Changes of DNAJB6a and DNAJB6b mRNA levels in CRC specimens were statistically analyzed using the GEO dataset.DLD-1 and HCT116 cells were treated with PI3K/mTOR dual inhibitor BEZ235 and control cells were treated with solvents.Then,transwell invasion and migration assays were performed and the number of transmembrane cells were counted.Constitutively activated AKT (myr-Akt) was transiently-transfected into DNAJB6b stably-depleted DLD-1 and HCT116 cells (Referred as DLD-1-shD6b,HCT116-shD6b in this paper) and the controls were transfected with the empty vector.The rescue assay were conducted,changes in protein expression levels were detected by Western blot,and levels of cell invasion and migration were evaluated by Transwell assays. RESULTS: Compared with adjacent normal tissues,mRNA expressions of DNAJB6b but not DNAJB6a were significantly up-regulated in the CRC tissues (P<0.05).DNAJB6b depletion markedly reduced the p-Akt (Ser473) protein levels in DLD-1 and HCT116 cells compared with the control groups.The numbers of transmembrane cells treated by BEZ235 were significantly reduced compared to only about 20% of the control group (P<0.01).Resultsfrom the rescue assay showed that enforced expression of exogenous myr-Akt in DLD-1-shD6b and HCT116-shD6b cells up-regulated the p-Akt (Ser473) levels and prominently reversed the decreased number of transmembrane cells caused by DNAJB6b knockdown (P<0.01). CONCLUSION: DNAJB6b was up-regulated in CRC tissues,and its overexpression enhanced the invasion and migration abilities of CRC cells by activating the AKT pathway.Our observations suggest that these abnormal expressions played an oncogenic role in the development of CRC.

Key words: colorectal cancer, DNAJB6b, invasion and migration, AKT, oncogenic role

CLC Number:

R73-37

CHEN Dingxiong, ZHU Yiqing, SHI Jianhong, CAI Yan, HAO Jiajie, WANG Mingrong, LIANG Jianwei, ZHANG Yu. DNAJB6b enhanced invasion and migration abilities of colorectal cancer cells by activating the AKT pathway[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2022, 34(3): 178-183.

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DNAJB6b enhanced invasion and migration abilities of colorectal cancer cells by activating the AKT pathway

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