Knockdown of Matrix Metallopeptidase 9 Inhibits Metastasis of Oral Squamous Cell Carcinoma Cells in a Zebrafish Xenograft Model

Abstract

Destruction of extracellular matrix (ECM) is one of the basic steps of tumor invasion and metastasis. Matrix metalloproteinase (MMP) 9, a kind of zinc-ion-dependent endopeptidase, can degrade almost all protein components in the ECM, destroy the histological barrier of tumor cell invasion, and play a key role in tumor invasion and metastasis. The role of MMP-9 in tumor invasion and metastasis has attracted increasing attention and is considered the main proteolytic enzyme in this process. Although the overexpression of MMP-9 was detected in Oral squamous cell carcinoma (OSCC) tissues, further basic studies in vivo and in vitro are needed to investigate the role of MMP-9 in OSCCs and provide scientific validation. In this research, we developed a novel OSCC zebrafish xenograft model to study the role of the MMP-9 gene in oral carcinogenesis. Firstly, the MMP-9/shRNA lentiviral clone and control virus were constructed and transfected into OSCC cells. Then, the decreasing expression of MMP-9 was verified by RT-PCR and immunocytochemistry. Cell proliferation was detected by MTT assay. Colony formation was evaluated by colony formation assay. Cell invasion was evaluated using transwell invasion assay in vitro. In addition, OSCC cells with MMP-9/shRNA knockdown and control vector were injected into zebrafish and an OSCC tumor model in zebrafish was established to evaluate invasion and metastasis in vivo. Knockdown of MMP-9 gene by shRNA could inhibit OSCC cell growth and clone formation and markedly suppress cell invasion in vitro. And the knockdown of the MMP-9 gene could also significantly decrease the metastatic distance and number of metastatic tumor cells or lesions in vivo and suppress the metastasis rate in xenografted zebrafish. Taken together, these evidences indicated that the knockdown of MMP-9 might suppress OSCC cell invasion and metastasis in vivo and in vitro. The MMP-9 gene may be a promising therapeutic target for OSCCs in the future.

MMP-9 Knockdown Inhibits Oral Squamous Cell Carcinoma Lymph Node Metastasis in the Nude Mouse Tongue-Xenografted Model through the RhoC/Src Pathway

“…All cells were incubated at 37°C with 5% CO 2 . The stably transfected OSCC cell lines CAL27/MMP-9/shRNA and SCC15/MMP-9/shRNA and their control cells transfected with the control vector were previously established based on HIV lentivirus transfection [9]. University.…”

“…Our preliminary data found that knockdown of MMP-9 could inhibit OSCC cell metastasis in the zebrafish model [ 9 ]. To further confirm the role of MMP-9 in metastasis and explore its potential underlying mechanism, we established a BALB/c nude mouse tongue orthotopic OSCC cell xenografted model.…”

Unveiling Tumor Microenvironment Interactions Using Zebrafish Models

“…In early invasion, remodeling of the ECM, for example, can be achieved through the release of MMPs by stromal, inflammatory, or cancer cells (Lu et al, 2011 ). Although clinical studies using a broad range of MMP inhibitors have historically been unsuccessful, the exploration into the roles of MMPs and the pathways regulating their expression and activation (especially of MMP2 and MMP9) continues onward with the aid of zebrafish xenografts (Wyatt et al, 2017 ; Simbulan-Rosenthal et al, 2019 ; Wen et al, 2020 ). Integrin proteins have also attracted many researchers' attention.…”

The SWI/SNF chromatin remodeling assemblies BAF and PBAF differentially regulate cell cycle exit and cellular invasion in vivo

“…A variety of in vivo invasion models have been studied, including cancer xenograft models in mouse (9)(10)(11) and zebrafish (12,13), which each have their respective benefits and drawbacks. Over the past ~15 years, Caenorhabditis elegans anchor cell (AC) invasion has emerged as a unique alternative model due to the visually tractable single-cell nature of its cell invasion (Fig 1A) (14).…”

“…While in vitro invasion models provide an efficient means to study the mechanical aspects of cellular invasion, they are currently unable to replicate the complex microenvironment in which cells must invade during animal development and disease. A variety of in vivo invasion models have been studied, including cancer xenograft models in mouse (9)(10)(11) and zebrafish (12,13), each having their respective benefits and drawbacks. Over the past ~15 years, Caenorhabditis elegans anchor cell (AC) invasion has emerged as a powerful alternative model due to its visually tractable single-cell nature (Fig 1 A) (14).…”