TIMP3 regulates osteosarcoma cell migration, invasion, and chemotherapeutic resistances

Abstract

Tissue inhibitors of metalloproteinases (TIMPs) inhibit matrix metalloproteinases (MMPs) to limit degradation of the extracellular matrix. Low levels of TIMP3 have been demonstrated in cancer tissues at advanced clinical stages, with positive distant metastasis and chemotherapeutic resistance. We examined the role of TIMP3 in osteosarcoma (OS) cell invasiveness and chemoresistance. TIMP3 was overexpressed or knocked down in the human OS cell lines Saos2 and MG63. Cell migration and invasion capacities were then evaluated using Transwell assays, and resistance to cisplatin was assessed by CCK-8 assay and flow cytometry. Real-time PCR and western blotting were used to investigate activation of signaling pathways downstream of TIMP3. Overexpression of TIMP3 inhibited the migration and invasion of Saos2 and MG63 cells, while knockdown of TIMP3 had the opposite effect. Cell survival after exposure to cisplatin was inhibited by TIMP3 overexpression in both Saos2 and MG63 cells. Consistently, downregulation of TIMP3 gene expression significantly decreased the sensitivity of OS cells to cisplatin treatment. MMP1, MMP2, Bcl-2, and Akt1 were all downregulated following TIMP3 overexpression, while Bax and cleaved caspase-3 were upregulated. TIMP3 knockdown had opposite effects on the regulation of these genes. Taken together, our findings suggest TIMP3 as a new target for inhibition of OS progression and chemotherapeutic resistance.

Suppression of human breast cancer cells by tectorigenin through downregulation of matrix metalloproteinases and MAPK signaling in�vitro

“…Apoptosis is an important mode of cell death during tumor chemotherapy. The caspase pathway is usually involved in the process of apoptosis, and CASP-3 is the main apoptogenic protein downstream of the mitochondrial apoptosis signaling pathways (17); it cleaves various key cellular substrates, therefore resulting in apoptosis (36). Cleavage of PARP is another defining characteristic of apoptosis, and serves a pivotal role in it (37).…”

“…Measurement of cell viability. Cell viability was measured using the Cell Counting kit-8 (CCK-8), as previously described (17). Briefly, MDA-MB-231 and MCF-7 cells (6,000 cells/well) were plated in a 96-well plate and incubated overnight at 37˚C without drug treatment.…”

TIMP3 promoter hypermethylation contributes to the risk of renal cell carcinoma: A meta-analysis

“…TIMP3 gene, found in chromosome 22q13.2, is a tumor suppressor gene. It is involved in the inhibition of the growth, angiogenesis, invasion, and metastasis of human cancers [24][25][26]. Decrease in TIMP3 expression is associated with abnormal promoter methylation in various malignant tumors, including kidney cancer [11,27], bladder cancer [28], choriocarcinoma [29,30], esophageal squamous cell carcinoma [31], meningioma [32], and gastric cancer [33].…”

Carthamin yellow inhibits matrix degradation and inflammation induced by LPS in the intervertebral disc via suppression of MAPK pathway activation

“…A cell counting kit-8 (CCK8) (Dojindo, Kumamoto, Japan) was used to test the viability of NP cells after CY treatment for 1 day according to previously reported methods (24). Approximately 5×10 3 NP cells were seeded on each film and transferred to 96-well plates.…”