Expression of TIMPs and MMPs in Ovarian Tumors, Ascites, Ascites-Derived Cells, and Cancer Cell Lines: Characteristic Modulatory Response Before and After Chemotherapy Treatment

Ruth M Escalona; George Kannourakis; Jock K Findlay; Nuzhat Ahmed

doi:10.3389/fonc.2021.796588

Expression of TIMPs and MMPs in Ovarian Tumors, Ascites, Ascites-Derived Cells, and Cancer Cell Lines: Characteristic Modulatory Response Before and After Chemotherapy Treatment

Front Oncol. 2022 Jan 3:11:796588. doi: 10.3389/fonc.2021.796588. eCollection 2021.

Authors

Ruth M Escalona^{1

2

3

4}, George Kannourakis^{1

5}, Jock K Findlay^{2

3

4}, Nuzhat Ahmed^{1

2

3

4

5}

Affiliations

¹ Fiona Elsey Cancer Research Institute, Ballarat, VIC, Australia.
² Department of Obstetrics and Gynaecology, University of Melbourne, Melbourne, VIC, Australia.
³ Centre for Reproductive Health, Hudson Institute of Medical Research, Melbourne, VIC, Australia.
⁴ Department of Translational Medicine, Monash University, Melbourne, VIC, Australia.
⁵ School of Science, Psychology and Sport, Federation University Australia, Ballarat, VIC, Australia.

Abstract

Background: The tissue inhibitors of metalloproteinase (TIMPs) and their associated metalloproteinase (MMPs) are essential regulators of tissue homeostasis and are essential for cancer progression. This study analyzed the expression of TIMP-1,-2,-3 and the associated MMPs (MMP-2,-9,-11,-14) in different Stages, Grades and World Health Organization (WHO) classifications of serous ovarian tumors, ascites, ascites-derived cells from chemo-naïve (CN) and relapsed (CR) patients, and in ovarian cancer cell lines. The status of TIMPs and associated MMPs in response to chemotherapy treatment was assessed in cancer cell lines; TCGA data was interrogated to gauge TIMPs and associated MMPs as prognostic and platinum-response indicators.

Methods: The levels of TIMP-1, -2 and -3 were assessed by immunohistochemistry. The mRNA expression of TIMPs and MMPs was quantified by real time PCR (qRT-PCR). The chemosensitivity (IC₅₀ values) to Cisplatin or Paclitaxel in cell lines was evaluated by MTT assay. The levels of TIMPs in ascites and cell lysates were analyzed by an ELISA assay.

Results: The expression of TIMP-2 was significantly upregulated in Type 2 compared to Type 1 tumors and normal/benign ovarian tissues. TIMP-3 expression was significantly enhanced in Stage III, Grade 3 and Type 2 tumors compared to normal/benign ovarian tissues. The mRNA expression of MMP-9,-11 and -14 was significantly upregulated in Stage IV compared to normal/benign ovarian tissues. The expression of TIMP-1 was highest, followed by TIMP-2 and then TIMP-3 in CN ascites. At the cellular level, TIMP-2 mRNA expression was significantly higher in CN compared to CR epithelial cells in patients. The expression of TIMP-1 and -2, MMPs and cancer stem cells (CSCs) were upregulated in response to chemotherapy treatments in cancer cell lines. Interrogation of the TCGA dataset suggests shifts in platinum responses in patients consistent with genetic alterations in TIMP-2, -3 and MMP-2, -11 genes in tumors; and decreased overall survival (OS) and progression-free survival (PFS) in patients with altered MMP-14 genes.

Conclusions: TIMPs and related MMPs are differentially expressed in serous ovarian tumors, ascites, ascites-derived cells and ovarian cancer cell lines. Chemotherapy treatment modulates expression of TIMPs and MMPs in association with increased expression of genes related to cancer stem cells.

Keywords: cancer stem cells (CSCs); chemo-naïve (CN); metalloproteinases (MMPs); platinum resistance; relapsed (CR); tissue inhibitors of metalloproteinases (TIMPs).