1. TROP-2 exhibits tumor suppressive functions in cervical cancer by dual inhibition of IGF-1R and ALK signaling 
Sarah T.K. Sin, Yan Li, Ming Liu, Stephanie Ma, Xin-Yuan Guan 
Gynecologic Oncology 
Volume 152, Issue 1, Pages (January 2019)
DOI: /j.ygyno
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2. Fig. 1
TROP-2 exhibits potent tumor suppressive functions in HeLa cells. (A) Western blotting showing overexpression of TROP-2 in HeLa cells. β-actin was used as loading control. (B) XTT assay detecting cell proliferation of vector control cells and TROP-2-overexpressing clones of HeLa cells. *P < 0.05, ***P < 0.001, Student's t-test. (C) Foci formation and soft agar assays of vector control and TROP-2-overexpressing clones of HeLa cells. ***P < 0.001, Student's t-test. (D) Migration and invasion assays of vector control and TROP-2-overexpressed clones of HeLa. **P < 0.01, ***P < 0.001, Student's t-test. (E) In vivo tumor formation assay of TROP-2-overexpressed and control cells of HeLa. ***P < 0.001, Student's t-test. (F) Lung metastasis of HeLa-Vec and HeLa-TROP2 (C9) cells injected through tail vain in SCID mice. Arrow heads indicate positions of tumor nodules; dotted lines indicate tumor nodule areas.
Gynecologic Oncology  , DOI: ( /j.ygyno )
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3. Fig. 2
Knockdown of TROP-2 promotes oncogenicity of HeLa cells. (A) Western blotting showing knockdown of TROP-2 in HeLa cells. β-actin was used as loading control. (B) XTT assay detecting cell proliferation of vector control and TROP-2 knockdown cells of HeLa. ***P < 0.001, Student's t-test. (C) Foci formation and soft agar assays of vector control and TROP-2 knockdown cells of HeLa. *P < 0.05, **P < 0.01, ***P < 0.001, Student's t-test. (D) Migration and invasion assays of vector control and TROP-2 knockdown cells of HeLa. **P < 0.01, ***P < 0.001, Student's t-test. (E) In vivo tumor formation assay of TROP-2 knockdown and control cells of HeLa (n = 5). *P < 0.05, Student's t-test.
Gynecologic Oncology  , DOI: ( /j.ygyno )
Copyright © 2018 The Authors Terms and Conditions

4. Fig. 3
TROP-2 promotes cisplatin-induced cell apoptosis. (A) Cell viability assay on TROP-2-overexpressed and control cells of HeLa after treatment of gradient concentrations of cisplatin. *P < 0.05, **P < 0.01, ***P < 0.001, Student's t-test. (B) Cell viability assay on TROP-2 knockdown and control cells of HeLa after treatment of gradient concentrations of cisplatin. *P < 0.05, **P < 0.01, ***P < 0.001, Student's t-test. (C) TUNEL assay on TROP-2-overexpressed and control cells of HeLa after cisplatin treatment (8 μg/ml, 24 h). ***P < 0.001, Student's t-test. (D) Western blot analysis of (cleaved) PARP with or without cisplatin treatment in HeLa-Vec and HeLa-TROP2 cells. β-actin was used as loading control.
Gynecologic Oncology  , DOI: ( /j.ygyno )
Copyright © 2018 The Authors Terms and Conditions

5. Fig. 4
TROP-2 down-regulation activates IGF-1R and ALK signaling pathways. (A) Human phospho-RTK array of HeLa-Vec and HeLa-sh2 cell lysates. Signal intensities at the same locations of two membranes indicate the expression levels of the same phospho-RTK in the two samples. (B) Confirmation of phospho-RTK array results using Western blotting. (p-)IGF-1R, (p-)ALK, (p-)AKT and (p-)STAT3 were detected on TROP-2-overexpressed and knockdown cells and the respective control cells of HeLa. β-actin was used as loading control. (C) Vector and TROP-2 expressing plasmids were transiently co-transfected with IGF-1 or MDK in HeLa cells. Immunoprecipitation and Western blotting were applied to detect the interactions between TROP-2 and IGF-1 or MDK. (D) The predicted binding free energies between TROP-2 and IGF-1 and between TROP-2 and MDK using PPA-Pred2 (Protein-Protein Affinity Predictor).
Gynecologic Oncology  , DOI: ( /j.ygyno )
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6. Fig. 5
TROP-2-overexpressed cells are relatively irresponsive to the inhibition of IGF-1R and ALK. (A) Comparison of cell viability between vector- and TROP-2-transfected HeLa cells after AG1024 treatment for 24 h. *P < 0.05, **P < 0.01, ***P < 0.001, Student's t-test. (B) Comparison of cell viability between vector- and TROP-2-transfected HeLa cells after Crizotinib treatment for 24 h. *P < 0.05, Student's t-test. (C) Comparison of cell migration capabilities between vector- and TROP-2-transfected HeLa cells when incubated with AG1024 or Crizotinib for 48 h. ***P < 0.001, Student's t-test.
Gynecologic Oncology  , DOI: ( /j.ygyno )
Copyright © 2018 The Authors Terms and Conditions