Pro-invasive Effect of Proto-oncogene PBF Is Modulated by an Interaction with Cortactin.
Metastatic disease is responsible for the majority of endocrine cancer deaths. New therapeutic targets are urgently needed to improve patient survival rates.



The proto-oncogene PTTG1-binding factor (PBF/PTTG1IP) is overexpressed in multiple endocrine cancers and circumstantially associated with tumor aggressiveness. This study aimed to understand the role of PBF in tumor cell invasion and identify possible routes to inhibit its action. Design, Setting, Patients, and Interventions: Thyroid, breast, and colorectal cells were transfected with PBF and cultured for in vitro analysis. PBF and cortactin (CTTN) expression was determined in differentiated thyroid cancer and The Cancer Genome Atlas RNA-seq data.



Pro-invasive effects of PBF were evaluated by 2D Boyden chamber, 3D organotypic, and proximity ligation assays.



Our study identified that PBF and CTTN physically interact and co-localize, and that this occurs at the cell periphery, particularly at the leading edge of migrating cancer cells. Critically, PBF induces potent cellular invasion and migration in thyroid and breast cancer cells, which is entirely abrogated in the absence of CTTN. Importantly, we found that CTTN is over-expressed in differentiated thyroid cancer, particularly in patients with regional lymph node metastasis, which significantly correlates with elevated PBF expression. Mutation of PBF (Y174A) or pharmacological intervention modulates the PBF: CTTN interaction and attenuates the invasive properties of cancer cells.



Our results demonstrate a unique role for PBF in regulating CTTN function to promote endocrine cell invasion and migration, as well as identify a new targetable interaction to block tumor cell movement.
Institute of Metabolism and Systems Research (R.J.W., W.I., M.L.R., N.S., V.L.P., S.B., R.F., H.R.N., K.B., V.E.S., C.J.M.), University of Birmingham, Birmingham B15 2TT, United Kingdom; Department of Surgery, Faculty of Medicine (W.I.), Siriraj Hospital, Mahidol University, Bangkok 10700, Thailand; Section of Endocrinology and Internal Medicine (E.G.), University of Ferrara, 44121 Ferrara, Italy; STIM Laboratory (E.B.), University of Poitiers, 86073 Poitiers Cedex 9, France; Northern Centre for Cancer Care (U.M.), Freeman Hospital, Newcastle upon Tyne NE7 7DN, United Kingdom; Cancer Research United Kingdom & UCL Cancer Trials Centre (A.H.), University College London, London WC1E 6BT, United Kingdom; and Institute of Cancer and Genomic Sciences (H.M.), University of Birmingham, Birmingham B15 2TT, United Kingdom.
Rachel J
RJ
Watkins
Institute of Metabolism and Systems Research (R.J.W., W.I., M.L.R., N.S., V.L.P., S.B., R.F., H.R.N., K.B., V.E.S., C.J.M.), University of Birmingham, Birmingham B15 2TT, United Kingdom; Department of Surgery, Faculty of Medicine (W.I.), Siriraj Hospital, Mahidol University, Bangkok 10700, Thailand; Section of Endocrinology and Internal Medicine (E.G.), University of Ferrara, 44121 Ferrara, Italy; STIM Laboratory (E.B.), University of Poitiers, 86073 Poitiers Cedex 9, France; Northern Centre for Cancer Care (U.M.), Freeman Hospital, Newcastle upon Tyne NE7 7DN, United Kingdom; Cancer Research United Kingdom & UCL Cancer Trials Centre (A.H.), University College London, London WC1E 6BT, United Kingdom; and Institute of Cancer and Genomic Sciences (H.M.), University of Birmingham, Birmingham B15 2TT, United Kingdom.
Waraporn
W
Imruetaicharoenchoke
Institute of Metabolism and Systems Research (R.J.W., W.I., M.L.R., N.S., V.L.P., S.B., R.F., H.R.N., K.B., V.E.S., C.J.M.), University of Birmingham, Birmingham B15 2TT, United Kingdom; Department of Surgery, Faculty of Medicine (W.I.), Siriraj Hospital, Mahidol University, Bangkok 10700, Thailand; Section of Endocrinology and Internal Medicine (E.G.), University of Ferrara, 44121 Ferrara, Italy; STIM Laboratory (E.B.), University of Poitiers, 86073 Poitiers Cedex 9, France; Northern Centre for Cancer Care (U.M.), Freeman Hospital, Newcastle upon Tyne NE7 7DN, United Kingdom; Cancer Research United Kingdom & UCL Cancer Trials Centre (A.H.), University College London, London WC1E 6BT, United Kingdom; and Institute of Cancer and Genomic Sciences (H.M.), University of Birmingham, Birmingham B15 2TT, United Kingdom.
Martin L
ML
Read
Institute of Metabolism and Systems Research (R.J.W., W.I., M.L.R., N.S., V.L.P., S.B., R.F., H.R.N., K.B., V.E.S., C.J.M.), University of Birmingham, Birmingham B15 2TT, United Kingdom; Department of Surgery, Faculty of Medicine (W.I.), Siriraj Hospital, Mahidol University, Bangkok 10700, Thailand; Section of Endocrinology and Internal Medicine (E.G.), University of Ferrara, 44121 Ferrara, Italy; STIM Laboratory (E.B.), University of Poitiers, 86073 Poitiers Cedex 9, France; Northern Centre for Cancer Care (U.M.), Freeman Hospital, Newcastle upon Tyne NE7 7DN, United Kingdom; Cancer Research United Kingdom & UCL Cancer Trials Centre (A.H.), University College London, London WC1E 6BT, United Kingdom; and Institute of Cancer and Genomic Sciences (H.M.), University of Birmingham, Birmingham B15 2TT, United Kingdom.
Neil
N
Sharma
Institute of Metabolism and Systems Research (R.J.W., W.I., M.L.R., N.S., V.L.P., S.B., R.F., H.R.N., K.B., V.E.S., C.J.M.), University of Birmingham, Birmingham B15 2TT, United Kingdom; Department of Surgery, Faculty of Medicine (W.I.), Siriraj Hospital, Mahidol University, Bangkok 10700, Thailand; Section of Endocrinology and Internal Medicine (E.G.), University of Ferrara, 44121 Ferrara, Italy; STIM Laboratory (E.B.), University of Poitiers, 86073 Poitiers Cedex 9, France; Northern Centre for Cancer Care (U.M.), Freeman Hospital, Newcastle upon Tyne NE7 7DN, United Kingdom; Cancer Research United Kingdom & UCL Cancer Trials Centre (A.H.), University College London, London WC1E 6BT, United Kingdom; and Institute of Cancer and Genomic Sciences (H.M.), University of Birmingham, Birmingham B15 2TT, United Kingdom.
Vikki L
VL
Poole
Institute of Metabolism and Systems Research (R.J.W., W.I., M.L.R., N.S., V.L.P., S.B., R.F., H.R.N., K.B., V.E.S., C.J.M.), University of Birmingham, Birmingham B15 2TT, United Kingdom; Department of Surgery, Faculty of Medicine (W.I.), Siriraj Hospital, Mahidol University, Bangkok 10700, Thailand; Section of Endocrinology and Internal Medicine (E.G.), University of Ferrara, 44121 Ferrara, Italy; STIM Laboratory (E.B.), University of Poitiers, 86073 Poitiers Cedex 9, France; Northern Centre for Cancer Care (U.M.), Freeman Hospital, Newcastle upon Tyne NE7 7DN, United Kingdom; Cancer Research United Kingdom & UCL Cancer Trials Centre (A.H.), University College London, London WC1E 6BT, United Kingdom; and Institute of Cancer and Genomic Sciences (H.M.), University of Birmingham, Birmingham B15 2TT, United Kingdom.
Erica
E
Gentilin
Institute of Metabolism and Systems Research (R.J.W., W.I., M.L.R., N.S., V.L.P., S.B., R.F., H.R.N., K.B., V.E.S., C.J.M.), University of Birmingham, Birmingham B15 2TT, United Kingdom; Department of Surgery, Faculty of Medicine (W.I.), Siriraj Hospital, Mahidol University, Bangkok 10700, Thailand; Section of Endocrinology and Internal Medicine (E.G.), University of Ferrara, 44121 Ferrara, Italy; STIM Laboratory (E.B.), University of Poitiers, 86073 Poitiers Cedex 9, France; Northern Centre for Cancer Care (U.M.), Freeman Hospital, Newcastle upon Tyne NE7 7DN, United Kingdom; Cancer Research United Kingdom & UCL Cancer Trials Centre (A.H.), University College London, London WC1E 6BT, United Kingdom; and Institute of Cancer and Genomic Sciences (H.M.), University of Birmingham, Birmingham B15 2TT, United Kingdom.
Sukhchain
S
Bansal
Institute of Metabolism and Systems Research (R.J.W., W.I., M.L.R., N.S., V.L.P., S.B., R.F., H.R.N., K.B., V.E.S., C.J.M.), University of Birmingham, Birmingham B15 2TT, United Kingdom; Department of Surgery, Faculty of Medicine (W.I.), Siriraj Hospital, Mahidol University, Bangkok 10700, Thailand; Section of Endocrinology and Internal Medicine (E.G.), University of Ferrara, 44121 Ferrara, Italy; STIM Laboratory (E.B.), University of Poitiers, 86073 Poitiers Cedex 9, France; Northern Centre for Cancer Care (U.M.), Freeman Hospital, Newcastle upon Tyne NE7 7DN, United Kingdom; Cancer Research United Kingdom & UCL Cancer Trials Centre (A.H.), University College London, London WC1E 6BT, United Kingdom; and Institute of Cancer and Genomic Sciences (H.M.), University of Birmingham, Birmingham B15 2TT, United Kingdom.
Emy
E
Bosseboeuf
Institute of Metabolism and Systems Research (R.J.W., W.I., M.L.R., N.S., V.L.P., S.B., R.F., H.R.N., K.B., V.E.S., C.J.M.), University of Birmingham, Birmingham B15 2TT, United Kingdom; Department of Surgery, Faculty of Medicine (W.I.), Siriraj Hospital, Mahidol University, Bangkok 10700, Thailand; Section of Endocrinology and Internal Medicine (E.G.), University of Ferrara, 44121 Ferrara, Italy; STIM Laboratory (E.B.), University of Poitiers, 86073 Poitiers Cedex 9, France; Northern Centre for Cancer Care (U.M.), Freeman Hospital, Newcastle upon Tyne NE7 7DN, United Kingdom; Cancer Research United Kingdom & UCL Cancer Trials Centre (A.H.), University College London, London WC1E 6BT, United Kingdom; and Institute of Cancer and Genomic Sciences (H.M.), University of Birmingham, Birmingham B15 2TT, United Kingdom.
Rachel
R
Fletcher
Institute of Metabolism and Systems Research (R.J.W., W.I., M.L.R., N.S., V.L.P., S.B., R.F., H.R.N., K.B., V.E.S., C.J.M.), University of Birmingham, Birmingham B15 2TT, United Kingdom; Department of Surgery, Faculty of Medicine (W.I.), Siriraj Hospital, Mahidol University, Bangkok 10700, Thailand; Section of Endocrinology and Internal Medicine (E.G.), University of Ferrara, 44121 Ferrara, Italy; STIM Laboratory (E.B.), University of Poitiers, 86073 Poitiers Cedex 9, France; Northern Centre for Cancer Care (U.M.), Freeman Hospital, Newcastle upon Tyne NE7 7DN, United Kingdom; Cancer Research United Kingdom & UCL Cancer Trials Centre (A.H.), University College London, London WC1E 6BT, United Kingdom; and Institute of Cancer and Genomic Sciences (H.M.), University of Birmingham, Birmingham B15 2TT, United Kingdom.
Hannah R
HR
Nieto
Institute of Metabolism and Systems Research (R.J.W., W.I., M.L.R., N.S., V.L.P., S.B., R.F., H.R.N., K.B., V.E.S., C.J.M.), University of Birmingham, Birmingham B15 2TT, United Kingdom; Department of Surgery, Faculty of Medicine (W.I.), Siriraj Hospital, Mahidol University, Bangkok 10700, Thailand; Section of Endocrinology and Internal Medicine (E.G.), University of Ferrara, 44121 Ferrara, Italy; STIM Laboratory (E.B.), University of Poitiers, 86073 Poitiers Cedex 9, France; Northern Centre for Cancer Care (U.M.), Freeman Hospital, Newcastle upon Tyne NE7 7DN, United Kingdom; Cancer Research United Kingdom & UCL Cancer Trials Centre (A.H.), University College London, London WC1E 6BT, United Kingdom; and Institute of Cancer and Genomic Sciences (H.M.), University of Birmingham, Birmingham B15 2TT, United Kingdom.
Ujjal
U
Mallick
Institute of Metabolism and Systems Research (R.J.W., W.I., M.L.R., N.S., V.L.P., S.B., R.F., H.R.N., K.B., V.E.S., C.J.M.), University of Birmingham, Birmingham B15 2TT, United Kingdom; Department of Surgery, Faculty of Medicine (W.I.), Siriraj Hospital, Mahidol University, Bangkok 10700, Thailand; Section of Endocrinology and Internal Medicine (E.G.), University of Ferrara, 44121 Ferrara, Italy; STIM Laboratory (E.B.), University of Poitiers, 86073 Poitiers Cedex 9, France; Northern Centre for Cancer Care (U.M.), Freeman Hospital, Newcastle upon Tyne NE7 7DN, United Kingdom; Cancer Research United Kingdom & UCL Cancer Trials Centre (A.H.), University College London, London WC1E 6BT, United Kingdom; and Institute of Cancer and Genomic Sciences (H.M.), University of Birmingham, Birmingham B15 2TT, United Kingdom.
Allan
A
Hackshaw
Institute of Metabolism and Systems Research (R.J.W., W.I., M.L.R., N.S., V.L.P., S.B., R.F., H.R.N., K.B., V.E.S., C.J.M.), University of Birmingham, Birmingham B15 2TT, United Kingdom; Department of Surgery, Faculty of Medicine (W.I.), Siriraj Hospital, Mahidol University, Bangkok 10700, Thailand; Section of Endocrinology and Internal Medicine (E.G.), University of Ferrara, 44121 Ferrara, Italy; STIM Laboratory (E.B.), University of Poitiers, 86073 Poitiers Cedex 9, France; Northern Centre for Cancer Care (U.M.), Freeman Hospital, Newcastle upon Tyne NE7 7DN, United Kingdom; Cancer Research United Kingdom & UCL Cancer Trials Centre (A.H.), University College London, London WC1E 6BT, United Kingdom; and Institute of Cancer and Genomic Sciences (H.M.), University of Birmingham, Birmingham B15 2TT, United Kingdom.
Hisham
H
Mehanna
Institute of Metabolism and Systems Research (R.J.W., W.I., M.L.R., N.S., V.L.P., S.B., R.F., H.R.N., K.B., V.E.S., C.J.M.), University of Birmingham, Birmingham B15 2TT, United Kingdom; Department of Surgery, Faculty of Medicine (W.I.), Siriraj Hospital, Mahidol University, Bangkok 10700, Thailand; Section of Endocrinology and Internal Medicine (E.G.), University of Ferrara, 44121 Ferrara, Italy; STIM Laboratory (E.B.), University of Poitiers, 86073 Poitiers Cedex 9, France; Northern Centre for Cancer Care (U.M.), Freeman Hospital, Newcastle upon Tyne NE7 7DN, United Kingdom; Cancer Research United Kingdom & UCL Cancer Trials Centre (A.H.), University College London, London WC1E 6BT, United Kingdom; and Institute of Cancer and Genomic Sciences (H.M.), University of Birmingham, Birmingham B15 2TT, United Kingdom.
Kristien
K
Boelaert
Institute of Metabolism and Systems Research (R.J.W., W.I., M.L.R., N.S., V.L.P., S.B., R.F., H.R.N., K.B., V.E.S., C.J.M.), University of Birmingham, Birmingham B15 2TT, United Kingdom; Department of Surgery, Faculty of Medicine (W.I.), Siriraj Hospital, Mahidol University, Bangkok 10700, Thailand; Section of Endocrinology and Internal Medicine (E.G.), University of Ferrara, 44121 Ferrara, Italy; STIM Laboratory (E.B.), University of Poitiers, 86073 Poitiers Cedex 9, France; Northern Centre for Cancer Care (U.M.), Freeman Hospital, Newcastle upon Tyne NE7 7DN, United Kingdom; Cancer Research United Kingdom & UCL Cancer Trials Centre (A.H.), University College London, London WC1E 6BT, United Kingdom; and Institute of Cancer and Genomic Sciences (H.M.), University of Birmingham, Birmingham B15 2TT, United Kingdom.
Vicki E
VE
Smith
Institute of Metabolism and Systems Research (R.J.W., W.I., M.L.R., N.S., V.L.P., S.B., R.F., H.R.N., K.B., V.E.S., C.J.M.), University of Birmingham, Birmingham B15 2TT, United Kingdom; Department of Surgery, Faculty of Medicine (W.I.), Siriraj Hospital, Mahidol University, Bangkok 10700, Thailand; Section of Endocrinology and Internal Medicine (E.G.), University of Ferrara, 44121 Ferrara, Italy; STIM Laboratory (E.B.), University of Poitiers, 86073 Poitiers Cedex 9, France; Northern Centre for Cancer Care (U.M.), Freeman Hospital, Newcastle upon Tyne NE7 7DN, United Kingdom; Cancer Research United Kingdom & UCL Cancer Trials Centre (A.H.), University College London, London WC1E 6BT, United Kingdom; and Institute of Cancer and Genomic Sciences (H.M.), University of Birmingham, Birmingham B15 2TT, United Kingdom.
Christopher J
CJ
McCabe
D001943:Breast Neoplasms / Q000378:metabolism*; D045744:Cell Line, Tumor; D015179:Colorectal Neoplasms / Q000378:metabolism*; D051356:Cortactin / Q000378:metabolism*; D005260:Female; D005786:Gene Expression Regulation; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins; D008565:Membrane Proteins / Q000378:metabolism*; D009361:Neoplasm Invasiveness; D000090063:Proto-Oncogene Mas; D013964:Thyroid Neoplasms / Q000378:metabolism
C495868:CTTN protein, human; D051356:Cortactin; D047908:Intracellular Signaling Peptides and Proteins; C000715470:MAS1 protein, human; D008565:Membrane Proteins; C112633:PTTG1IP protein, human; D000090063:Proto-Oncogene Mas