A Novel Cell Line Derived from Pleomorphic Adenoma Expresses MMP2, MMP9, TIMP1, TIMP2, and Shows Numeric Chromosomal Anomalies

Abstract

Pleomorphic adenoma is the most common salivary gland neoplasm, and it can be locally invasive, despite its slow growth. This study aimed to establish a novel cell line (AP-1) derived from a human pleomorphic adenoma sample to better understand local invasiveness of this tumor. AP-1 cell line was characterized by cell growth analysis, expression of epithelial and myoepithelial markers by immunofluorescence, electron microscopy, 3D cell culture assays, cytogenetic features and transcriptomic study. Expression of matrix metalloproteinases (MMPs) and their tissue inhibitors (TIMPs) was also analyzed by immunofluorescence and zymography. Furthermore, epithelial and myoepithelial markers, MMPs and TIMPs were studied in the tumor that originated the cell line. AP-1 cells showed neoplastic epithelial and myoepithelial markers, such as cytokeratins, vimentin, S100 protein and smooth-muscle actin. These molecules were also found in vivo, in the tumor that originated the cell line. MMPs and TIMPs were observed in vivo and in AP-1 cells. Growth curve showed that AP-1 exhibited a doubling time of 3.342 days. AP-1 cells grown inside Matrigel recapitulated tumor architecture. Different numerical and structural chromosomal anomalies were visualized in cytogenetic analysis. Transcriptomic analysis addressed expression of 7 target genes (VIM, TIMP2, MMP2, MMP9, TIMP1, ACTA2 e PLAG1). Results were compared to transcriptomic profile of non-neoplastic salivary gland cells (HSG). Only MMP9 was not expressed in both libraries, and VIM was expressed solely in AP-1 library. The major difference regarding gene expression level between AP-1 and HSG samples occurred for MMP2. This gene was 184 times more expressed in AP-1 cells. Our findings suggest that AP-1 cell line could be a useful model for further studies on pleomorphic adenoma biology.

miRNA expression profile of mucoepidermoid carcinoma

“…To confirm our results, we validated the miRNA expression results in two salivary gland cell lines: an epidermoid carcinoma cell line (A‐253) and a human salivary gland cell line (HSG) (Falcão et al., ) using qRT‐PCR. miR‐302a expression levels were elevated in A‐253 cells compared with that in HSG cells.…”

<p>The role of circadian clock genes in tumors</p>

“…Tumorigenesis is a very complex and diversified process. In addition to producing abnormal changes in the cell cycle, it also needs to inhibit the apoptotic signals generated via cell contact, develop new blood vessels to provide nutrition and oxygen to support tumor growth, and induce the capacity to migrate and invade 71,111–113. To date, many crucial genes involved in the cancer-related regulation of cell proliferation, apoptosis, invasion, metastasis, and angiogenesis have been documented to have periodic patterns of expression over a 24-h cycle, such as the proliferation gene Ki-67,114,115 the proto-oncogene Mdm2,52,89 the proapoptotic gene Bax, the antiapoptotic gene Bcl-2,111 the invasion and metastasis gene MMP9,116,117 and the angiogenic gene Vegf 112,114,118.…”

La celularidad y su relación con las características capsulares como factor influyente en las estrategias operatorias del adenoma pleomorfo

“…Overexpression of MMP leads to excessive breakdown of the extracellular matrix. This occurs in many pathological conditions, such as cancer invasion and metastasis, arteriosclerosis or angiogenesis, and fibrosis [8][9][10] . However, some studies have revealed that TIMPs may increase the probability of malignant changes 11,12 .…”