Opposing Roles of IGFBP-3 and Heparanase in Regulating A549 Lung Cancer Cell Survival
In this study, we examined the roles of heparanase and IGFBP-3 in regulating A549 and H1299 non-small-cell lung cancer (NSCLC) survival. We found that H1299 cells, known to be p53-null with no expression of IGFBP-3, had higher heparanase levels and activity and higher levels of heparan sulfate (HS)...
| Main Authors: | , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2022-11-01
|
| Series: | Cells |
| Subjects: | |
| Online Access: | https://www.mdpi.com/2073-4409/11/22/3533 |
| _version_ | 1827644789473935360 |
|---|---|
| author | Hind Al Khashali Jadziah Wareham Ravel Ray Ben Haddad Kai-Ling Coleman Robert Ranzenberger Patrick McCombs Jeffrey Guthrie Deborah Heyl Hedeel Guy Evans |
| author_facet | Hind Al Khashali Jadziah Wareham Ravel Ray Ben Haddad Kai-Ling Coleman Robert Ranzenberger Patrick McCombs Jeffrey Guthrie Deborah Heyl Hedeel Guy Evans |
| author_sort | Hind Al Khashali |
| collection | DOAJ |
| description | In this study, we examined the roles of heparanase and IGFBP-3 in regulating A549 and H1299 non-small-cell lung cancer (NSCLC) survival. We found that H1299 cells, known to be p53-null with no expression of IGFBP-3, had higher heparanase levels and activity and higher levels of heparan sulfate (HS) in the media compared to the media of A549 cells. Inhibiting heparanase activity or its expression using siRNA had no effect on the levels of IGFBP-3 in the media of A549 cells, reduced the levels of soluble HS fragments, and led to decreased interactions between IGFBP-3 and HS in the media. HS competed with HA for binding to IGFBP-3 or IGFBP-3 peptide (<sup>215</sup>-KKGFYKKKQCRPSKGRKR-<sup>232</sup>) but not the mutant peptide (K228AR230A). HS abolished the cytotoxic effects of IGFBP-3 but not upon blocking HA–CD44 signaling with the anti-CD44 antibody (5F12). Blocking HA–CD44 signaling decreased the levels of heparanase in the media of both A549 and H1299 cell lines and increased p53 activity and the levels of IGFBP-3 in A549 cell media. Knockdown of p53 led to increased heparanase levels and reduced IGFBP-3 levels in A549 cell media while knockdown of IGFBP-3 in A549 cells blocked p53 activity and increased heparanase levels in the media. |
| first_indexed | 2024-03-09T18:25:20Z |
| format | Article |
| id | doaj.art-5e5b41d03c05467f8e94d34253b7d251 |
| institution | Directory Open Access Journal |
| issn | 2073-4409 |
| language | English |
| last_indexed | 2024-03-09T18:25:20Z |
| publishDate | 2022-11-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Cells |
| spelling | doaj.art-5e5b41d03c05467f8e94d34253b7d2512023-11-24T07:56:59ZengMDPI AGCells2073-44092022-11-011122353310.3390/cells11223533Opposing Roles of IGFBP-3 and Heparanase in Regulating A549 Lung Cancer Cell SurvivalHind Al Khashali0Jadziah Wareham1Ravel Ray2Ben Haddad3Kai-Ling Coleman4Robert Ranzenberger5Patrick McCombs6Jeffrey Guthrie7Deborah Heyl8Hedeel Guy Evans9Chemistry Department, Eastern Michigan University, Ypsilanti, MI 48197, USAChemistry Department, Eastern Michigan University, Ypsilanti, MI 48197, USAChemistry Department, Eastern Michigan University, Ypsilanti, MI 48197, USAChemistry Department, Eastern Michigan University, Ypsilanti, MI 48197, USAChemistry Department, Eastern Michigan University, Ypsilanti, MI 48197, USAChemistry Department, Eastern Michigan University, Ypsilanti, MI 48197, USAChemistry Department, Eastern Michigan University, Ypsilanti, MI 48197, USAChemistry Department, Eastern Michigan University, Ypsilanti, MI 48197, USAChemistry Department, Eastern Michigan University, Ypsilanti, MI 48197, USAChemistry Department, Eastern Michigan University, Ypsilanti, MI 48197, USAIn this study, we examined the roles of heparanase and IGFBP-3 in regulating A549 and H1299 non-small-cell lung cancer (NSCLC) survival. We found that H1299 cells, known to be p53-null with no expression of IGFBP-3, had higher heparanase levels and activity and higher levels of heparan sulfate (HS) in the media compared to the media of A549 cells. Inhibiting heparanase activity or its expression using siRNA had no effect on the levels of IGFBP-3 in the media of A549 cells, reduced the levels of soluble HS fragments, and led to decreased interactions between IGFBP-3 and HS in the media. HS competed with HA for binding to IGFBP-3 or IGFBP-3 peptide (<sup>215</sup>-KKGFYKKKQCRPSKGRKR-<sup>232</sup>) but not the mutant peptide (K228AR230A). HS abolished the cytotoxic effects of IGFBP-3 but not upon blocking HA–CD44 signaling with the anti-CD44 antibody (5F12). Blocking HA–CD44 signaling decreased the levels of heparanase in the media of both A549 and H1299 cell lines and increased p53 activity and the levels of IGFBP-3 in A549 cell media. Knockdown of p53 led to increased heparanase levels and reduced IGFBP-3 levels in A549 cell media while knockdown of IGFBP-3 in A549 cells blocked p53 activity and increased heparanase levels in the media.https://www.mdpi.com/2073-4409/11/22/3533heparanaseIGFBP-3lung cancerheparan sulfatehyaluronanCD44 |
| spellingShingle | Hind Al Khashali Jadziah Wareham Ravel Ray Ben Haddad Kai-Ling Coleman Robert Ranzenberger Patrick McCombs Jeffrey Guthrie Deborah Heyl Hedeel Guy Evans Opposing Roles of IGFBP-3 and Heparanase in Regulating A549 Lung Cancer Cell Survival Cells heparanase IGFBP-3 lung cancer heparan sulfate hyaluronan CD44 |
| title | Opposing Roles of IGFBP-3 and Heparanase in Regulating A549 Lung Cancer Cell Survival |
| title_full | Opposing Roles of IGFBP-3 and Heparanase in Regulating A549 Lung Cancer Cell Survival |
| title_fullStr | Opposing Roles of IGFBP-3 and Heparanase in Regulating A549 Lung Cancer Cell Survival |
| title_full_unstemmed | Opposing Roles of IGFBP-3 and Heparanase in Regulating A549 Lung Cancer Cell Survival |
| title_short | Opposing Roles of IGFBP-3 and Heparanase in Regulating A549 Lung Cancer Cell Survival |
| title_sort | opposing roles of igfbp 3 and heparanase in regulating a549 lung cancer cell survival |
| topic | heparanase IGFBP-3 lung cancer heparan sulfate hyaluronan CD44 |
| url | https://www.mdpi.com/2073-4409/11/22/3533 |
| work_keys_str_mv | AT hindalkhashali opposingrolesofigfbp3andheparanaseinregulatinga549lungcancercellsurvival AT jadziahwareham opposingrolesofigfbp3andheparanaseinregulatinga549lungcancercellsurvival AT ravelray opposingrolesofigfbp3andheparanaseinregulatinga549lungcancercellsurvival AT benhaddad opposingrolesofigfbp3andheparanaseinregulatinga549lungcancercellsurvival AT kailingcoleman opposingrolesofigfbp3andheparanaseinregulatinga549lungcancercellsurvival AT robertranzenberger opposingrolesofigfbp3andheparanaseinregulatinga549lungcancercellsurvival AT patrickmccombs opposingrolesofigfbp3andheparanaseinregulatinga549lungcancercellsurvival AT jeffreyguthrie opposingrolesofigfbp3andheparanaseinregulatinga549lungcancercellsurvival AT deborahheyl opposingrolesofigfbp3andheparanaseinregulatinga549lungcancercellsurvival AT hedeelguyevans opposingrolesofigfbp3andheparanaseinregulatinga549lungcancercellsurvival |