Knockdown of KLK12 inhibits viability and induces apoptosis in human colorectal cancer HT-29 cell line

Li,Qianyuan; Zhou,Xiukou; Fang,Zhengyu; Zhou,Huamiao

doi:10.3892/ijmm.2019.4327

Knockdown of KLK12 inhibits viability and induces apoptosis in human colorectal cancer HT-29 cell line

Authors:
- Qianyuan Li
- Xiukou Zhou
- Zhengyu Fang
- Huamiao Zhou
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Affiliations: Department of Anorectal Surgery, The First Affiliated Hospital of Zhejiang Chinese Medical University, Hangzhou, Zhejiang 310006, P.R. China, Department of Oncology, The First Affiliated Hospital of Zhejiang Chinese Medical University, Hangzhou, Zhejiang 310006, P.R. China
Published online on: August 30, 2019 https://doi.org/10.3892/ijmm.2019.4327
Pages: 1667-1676
Copyright: © Li et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Kallikrein‑related peptidase 12 (KLK12) is overexpressed in cancer tissues including gastric, breast and prostate cancer. However, the role of KLK12 in colorectal cancer is not fully understood. In the present study, the level of KLK12 was determined by performing reverse transcription‑polymerase chain reaction (RT‑qPCR) in colorectal cancer tissues and cell lines. Lipofectamine® 2000 was used to transfect HT‑29 cells to overexpress and knockdown KLK12. Cell viability, migration, invasion and apoptosis were detected by MTT, wound healing, Transwell and flow cytometry assays, respectively. The mRNA and protein expression levels of EMT‑associated proteins, apoptosis‑associated proteins, phosphorylated adenosine monophosphate‑activated protein kinase (p‑AMPK) and phosphorylated mammalian target of rapamycin (p‑mTOR) were determined by RT‑qPCR and western blot analysis. It was identified that the KLK12 mRNA levels were increased significantly in colorectal cancer tissues and cell lines. KLK12 small interfering RNA inhibited cell viability, migration and invasion. Furthermore, epithelial‑mesenchymal transition (EMT)‑associated proteins were altered by siKLK12. Cell apoptosis was induced by KLK12 downregulation, which was demonstrated by the changes in apoptosis‑associated proteins; however, KLK12 overexpression produced the opposite effect. SiKLK12 enhanced the expression of p‑AMPK and suppressed the expression of p‑mTOR, while KLK12 overexpression had the opposite effect. Promotion of KLK12 overexpression‑induced cell viability was reversed by 5‑aminoimidazole‑4‑carboxamide ribonucleotide, an activator of the AMPK signaling pathway, and rapamycin, a specific inhibitor of the mTOR signaling pathway. Taken together, the results of the present study indicated that KLK12 was overexpressed in colorectal cancer and may regulate cell behavior, potentially via the AMPK and mTOR pathways.

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