The 3q Oncogene SEC62 Predicts Response to Neoadjuvant Chemotherapy and Regulates Tumor Cell Migration in Triple Negative Breast Cancer
Abstract
:1. Introduction
2. Results
2.1. Frequency of SEC62 Alterations in TCGA Data
2.2. Immunohistochemical Analysis of SEC62 Expression in TNBC Tissue Samples
2.3. Correlation of Patient Survival with SEC62 Expression
2.4. Effect of SEC62 Expression on CAL-120 (TNBC) Cell Proliferation and Migratory Potential
3. Discussion
4. Materials and Methods
4.1. Analysis of The Cancer Genome Atlas (TCGA) Data
4.2. Patients and Tissue Samples
4.3. IHC Analysis of Sec62
4.4. Cell Biological Experiments
4.5. Cell Culture and Transfection
4.6. Real-Time Cell Proliferation Analysis
4.7. Migration Potential Analysis
4.8. Western Blot Analyses
4.9. Statistical Analysis
5. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
- Sidoni, A.; Cavaliere, A.; Bellezza, G.; Scheibel, M.; Bucciarelli, E.J.T.B. Breast cancer in young women: Clinicopathological features and biological specificity. Breast 2003, 12, 247–250. [Google Scholar] [CrossRef] [PubMed]
- Radosa, J.C.; Eaton, A.; Stempel, M.; Khander, A.; Liedtke, C.; Solomayer, E.F.; Karsten, M.; Pilewskie, M.; Morrow, M.; King, T.A. Evaluation of Local and Distant Recurrence Patterns in Patients with Triple-Negative Breast Cancer According to Age. Ann. Surg. Oncol. 2017, 24, 698–704. [Google Scholar] [CrossRef] [PubMed]
- Collins, L.C.; Marotti, J.D.; Gelber, S.; Cole, K.; Ruddy, K.; Kereakoglow, S.; Brachtel, E.F.; Schapira, L.; Come, S.E.; Winer, E.P.; et al. Pathologic features and molecular phenotype by patient age in a large cohort of young women with breast cancer. Breast Cancer Res. Treat. 2012, 131, 1061–1066. [Google Scholar] [CrossRef] [PubMed]
- Xiong, Q.; Valero, V.; Kau, V.; Kau, S.W.; Taylor, S.; Smith, T.L.; Buzdar, A.U.; Hortobagyi, G.N.; Theriault, R.L. Female patients with breast carcinoma age 30 years and younger have a poor prognosis: The M.D. Anderson Cancer Center experience. Cancer 2001, 92, 2523–2528. [Google Scholar] [CrossRef] [PubMed]
- Untch, M.; Jackisch, C.; Schneeweiss, A.; Conrad, B.; Aktas, B.; Denkert, C.; Eidtmann, H.; Wiebringhaus, H.; Kümmel, S.; Hilfrich, J.; et al. Nab-paclitaxel versus solvent-based paclitaxel in neoadjuvant chemotherapy for early breast cancer (GeparSepto-GBG 69): A randomised, phase 3 trial. Lancet Oncol. 2016, 17, 345–356. [Google Scholar] [CrossRef]
- Gröbner, S.N.; Worst, B.C.; Weischenfeldt, J.; Buchhalter, I.; Kleinheinz, K.; Rudneva, V.A.; Johann, P.D.; Balasubramanian, G.P.; Segura-Wang, M.; Brabetz, S.; et al. The landscape of genomic alterations across childhood cancers. Nature 2018, 15, 321–327. [Google Scholar] [CrossRef]
- Cancer Genome Atlas Research Network. Comprehensive genomic characterization defines human glioblastoma genes and core pathways. Nature 2008, 23, 1061–1068. [Google Scholar] [CrossRef]
- Hagerstrand, D.; Tong, A.; Schumacher, S.E.; Ilic, N.; Shen, R.R.; Cheung, H.W.; Vazquez, F.; Shrestha, Y.; Kim, S.Y.; Giacomelli, A.O.; et al. Systematic interrogation of 3q26 identifies TLOC1 and SKL as cancer drivers. Cancer Discov. 2013, 3, 1044–1057. [Google Scholar] [CrossRef]
- Radosa, J.C.; Kasoha, M.; Schilz, A.-C.; Linxweiler, M.; Linxweiler, B.; Takacs, Z.F.; Kaya, A.; Bohle, R.M.; Wagner, M.; Wagenpfeil, G.; et al. Effect of the 3q26-coding oncogene SEC62 as a prognostic marker in patients with ovarian neoplasia. Front. Physiol. 2022, 13, 925023. [Google Scholar] [CrossRef]
- Woenckhaus, J.; Steger, K.; Werner, E.; Fenic, I.; Gamerdinger, U.; Dreyer, T.; Stahl, U. Genomic gain of PIK3CA and increased expression of p110α are associated with progression of dysplasia into invasive squamous cell carcinoma. J. Pathol. 2002, 198, 335–342. [Google Scholar] [CrossRef]
- Comtesse, N.; Keller, A.; Diesinger, I.; Bauer, C.; Kayser, K.; Huwer, H.; Lenhof, H.P.; Meese, E. Frequent overexpression of the genes FXR1, CLAPM1 and EIF4G located on amplicon 3q26-27 in squamous cell carcinoma of the lung. Int. J. Cancer 2007, 120, 2538–2544. [Google Scholar] [CrossRef]
- Zimmermann, J.S.M.; Linxweiler, J.; Radosa, J.C.; Linxweiler, M.; Zimmermann, R. The ER membrane protein Sec62 as potential therapeutic target in SEC62 overexpressing tumors. Front. Physiol. 2022, 13, 1014271. [Google Scholar] [CrossRef]
- Linxweiler, M.; Schorr, S.; Jung, M.; Schäuble, N.; Linxweiler, J.; Langer, F.; Schäfers, H.-J.; Cavalié, A.; Zimmermann, R.; Greiner, M. Targeting cell migration and the ER stress response with calmodulin antagonists: A clinically tested small molecule phenocopy of SEC62 gene silencing in human tumor cells. BMC Cancer 2013, 13, 574. [Google Scholar] [CrossRef]
- Linxweiler, M.; Linxweiler, J.; Barth, M.; Benedix, J.; Jung, V.; Kim, Y.-J.; Bohle, R.; Zimmermann, R.; Greiner, M. Sec62 bridges the gap from 3q amplification to molecular cell biology in Non-Small Cell Lung Cancer. Am. J. Pathol. 2012, 180, 473–483. [Google Scholar] [CrossRef]
- Linxweiler, M.; Bochen, F.; Schick, B.; Wemmert, S.; Al Kadah, B.; Greiner, M.; Hasenfus, A.; Bohle, R.-M.; Juhasz-Bӧss, I.; Solomayer, E.-F.; et al. Identification of SEC62 as a potential marker for 3q amplification and cellular migration in dysplastic cervical lesions. BMC Cancer 2016, 16, 676. [Google Scholar] [CrossRef]
- Wemmert, S.; Lindner, Y.; Linxweiler, J.; Wagenpfeil, S.; Bohle, R.; Niewald, M.; Schick, B. Initial evidence for Sec62 as a prognostic marker in advanced head and neck squamous cell carcinoma. Oncol. Lett. 2016, 11, 1661–1670. [Google Scholar] [CrossRef]
- Greiner, M.; Kreutzer, B.; Jung, V.; Grobholz, R.; Hasenfus, A.; Stöhr, R.; Franz, R.; Tornillo, L.; Dudek, J.; Stöckle, M.; et al. Silencing of the SEC62 gene inhibits migratory and invasive potential of various tumor cells. Int. J. Cancer 2011, 128, 2284–2295. [Google Scholar] [CrossRef]
- Hanahan, D.; Weinberg, R.A. The hallmarks of cancer. Cell 2000, 100, 57–70. [Google Scholar] [CrossRef]
- Takacs, F.Z.; Radosa, J.C.; Linxweiler, M.; Kasohah, M.; Bohle, R.M.; Bochen, F.; Unger, C.; Solomayer, E.-F.; Schick, B.; Juhasz-Böss, I. Identification of 3q oncogene SEC62 as a marker for distant metastasis and poor clinical outcome in invasive ductal breast cancer. Arch. Gynecol. Obstet. 2019, 299, 1405–1413. [Google Scholar] [CrossRef]
- Sinn, H.P.; Schmid, H.; Junkermann, H.; Huober, J.; Leppien, G.; Kaufmann, M.; Bastert, G.; Otto, H.F. Histologische Regression des Mammakarzinoms nach primärer (neoadjuvanter) Chemotherapie [Histologic regression of breast cancer after primary (neoadjuvant) chemotherapy]. Geburtshilfe Frauenheilkd. 1994, 54, 552–558. [Google Scholar] [CrossRef]
- Matuschek, C.; Jazmati, D.; Bölke, E.; Tamaskovics, B.; Corradini, S.; Budach, W.; Krug, D.; Mohrmann, S.; Ruckhäberle, E.; Fehm, T.; et al. Post-Neoadjuvant Treatment Strategies in Breast Cancer. Cancers 2022, 14, 1246. [Google Scholar] [CrossRef] [PubMed]
- Tutt, A.N.J.; Garber, J.E.; Kaufman, B.; Viale, G.; Fumagalli, D.; Rastogi, P.; Gelber, R.D.; de Azambuja, E.; Fielding, A.; Balmaña, J.; et al. Adjuvant Olaparib for Patients with BRCA1- or BRCA2-Mutated Breast Cancer. N. Engl. J. Med. 2021, 24, 2394–2405. [Google Scholar] [CrossRef] [PubMed]
- Liu, X.; Su, K.; Sun, X.; Jiang, Y.; Wang, L.; Hu, C.; Zhang, C.; Lu, M.; Du, X.; Xing, B. Sec62 promotes stemness and chemoresistance of human colorectal cancer through activating Wnt/β-catenin pathway. J. Exp. Clin. Cancer Res. 2021, 40, 132. [Google Scholar] [CrossRef] [PubMed]
- Jung, V.; Kamradt, J.; Kindich, R.; Jung, M.; Mueller, M.; Schulz, W.A.; Engers, R.; Stoeckle, M.; Zimmermann, R.; Wullich, B. Genomic and expression analysis of the 3q25-q26 amplicon reveals TLOC1/SEC62 as a probable target gene in prostate cancer. Mol. Cancer Res. 2006, 4, 169–176. [Google Scholar] [CrossRef]
- Weng, L.; Du, J.; Zhou, Q.; Cheng, B.; Li, J.; Zhang, D.; Ling, C. Identification of cyclin B1 and Sec62 as biomarkers for recurrence in patients with HBV-related hepatocellular carcinoma after surgical resection. Mol. Cancer 2012, 11, 39. [Google Scholar] [CrossRef]
- Bochen, F.; Adisurya, H.; Wemmert, S.; Lerner, C.; Greiner, M.; Zimmermann, R.; Hasenfus, A.; Wagner, M.; Smola, S.; Pfuhl, T.; et al. Effect of 3q oncogenes SEC62 and SOX2 on lymphatic metastasis and clinical outcome of head and neck squamous cell carcinomas. Oncotarget 2017, 8, 4922–4934. [Google Scholar] [CrossRef]
- Lakkaraju, A.K.K.; Thankappan, R.; Mary, C.; Garrison, J.L.; Taunton, J.; Strub, K. Efficient secretion of small proteins in mammalian cells relies on Sec62-dependent posttranslational translocation. Mol. Biol. Cell 2012, 23, 2712–2722. [Google Scholar] [CrossRef]
- Fumagalli, F.; Noack, J.; Bergmann, T.; Cebollero, E.; Pisoni, G.B.; Fasana, E.; Fregno, I.; Galli, C.; Loi, M.; Soldá, T.; et al. Translocon component Sec62 acts in endoplasmic reticulum turnover during stress recovery. Nat. Cell Biol. 2016, 18, 1173–1184. [Google Scholar] [CrossRef]
- Lang, S.; Benedix, J.; Fedeles, S.V.; Schorr, S.; Schirra, C.; Schäuble, N.; Jalal, C.; Greiner, M.; Haßdenteufel, S.; Tatzelt, J.; et al. Different effects of Sec61, Sec62 and Sec63-depletion on transport of polypeptides into the endoplasmic reticulum of mammalian cells. J. Cell Sci. 2012, 125, 1958–1969. [Google Scholar] [CrossRef]
- Carpenter, W.T., Jr.; Davis, J.M. Another view of the history of antipsychotic drug discovery and development. Mol. Psychiatry 2012, 17, 1168–1173. [Google Scholar] [CrossRef]
- Mahalingam, D.; Wilding, G.; Denmeade, S.; Sarantopoulas, J.; Cosgrove, D.; Cetnar, J.; Azad, N.; Bruce, J.; Kurman, M.; Allgood, V.E.; et al. Mipsagargin, a novel thapsigargin-based PSMA-activated prodrug: Results of a first-in-man phase I clinical trial in patients with refractory, advanced or metastatic solid tumours. Br. J. Cancer 2016, 114, 986–994. [Google Scholar] [CrossRef]
- Goldhirsch, A.; Winer, E.P.; Coates, A.S.; Gelber, R.D.; Piccart-Gebhart, M.; Thürlimann, B.; Senn, H.J. Personalizing the treatment of women with early breast cancer: Highlights of the St Gallen International Expert Consensus on the Primary Therapy of Early Breast Cancer 2013. Ann. Oncol. 2013, 24, 2206–2223. [Google Scholar] [CrossRef]
- Remmele, W.; Stegner, H.E. Vorschlag zur einheitlichen Definition eines Immunreaktiven Score (IRS) für den immunhistochemischen Ostrogenrezeptor-Nachweis (ER-ICA) im Mammakarzinomgewebe [Recommendation for uniform definition of an immunoreactive score (IRS) for immunohistochemical estrogen receptor detection (ER-ICA) in breast cancer tissue]. Pathologe 1987, 8, 138–140. (In German) [Google Scholar]
TNBC (n = 64) | |
---|---|
n (%) | |
Age (median (range)) | 55 (31–78) |
Menopausal status | |
Premenopausal | 21 (32.8%) |
Postmenopausal | 43 (67.2%) |
cT-stage | |
T1 | 22 (34.4%) |
T2 | 30 (46.9%) |
T3 | 2 (3.1%) |
T4 | 10 (15.6%) |
ypT-stage | |
T0 | 27 (42.2%) |
T1 | 24 (37.5%) |
T2 | 6 (9.4%) |
T3 | 5 (7.8%) |
T4 | 2 (3.1%) |
cN-stage | |
N0 | 31 (48.4%) |
N1 | 20 (31.3%) |
N2 | 6 (9.4%) |
N3 | 7 (10.9%) |
ypN-stage | |
N0 | 44 (68.8%) |
N1 | 10 (15.6%) |
N2 | 7 (10.9%) |
N3 | 3 (4.7%) |
Grading | |
G1 | 0 (0%) |
G2 | 12 (18.7%) |
G3 | 52 (81.3%) |
Ki67 | |
<15% | 0 (0%) |
≥15% | 64 (100%) |
Regression grade according to Sinn (median (range)) | 2 (0–4) |
Local recurrence | |
Yes | 4 (6.6%) |
No | 60 (93.4%) |
Distant metastasis | |
Yes | 25 (39.1%) |
No | 39 (60.9%) |
TNBC (n = 64) | p | |
---|---|---|
Median (Range) | ||
IRS core biopsy | 8 (2–12) | |
IRS final specimen | 4 (0–12) | ≤0.01 |
Difference between IRS core biopsy and IRS final specimen | 4 (−6–12) | |
n (%) | ||
IRS core biopsy | ||
negative | 3 (4.7%) | |
low | 8 (12.5%) | |
moderate | 32 (50%) | |
high | 21 (32.8%) | |
IRS final specimen | ||
negative | 28 (44.4%) | |
low | 11 (17.5%) | |
moderate | 15 (23.8%) | |
high | 9 (14.3%) | |
IRS ≤ 8 core biopsy | 42 (65.6%) | |
IRS > 8 core biopsy | 22 (34.4%) | |
IRS ≤ 8 final specimen | 51 (81%) | |
IRS > 8 final specimen | 12 (19%) | |
Difference between IRS core biopsy and IRS final specimen < 6 | 39 (61.9%) | |
Difference between IRS core biopsyand IRS final specimen ≥ 6 | 24 (38.1%) |
IRS ≤ 8 Core Biopsy n = 42 | IRS > 8 Core Biopsy n = 22 | p | |
Regression grade according to Sinn (median, range) | 2 (0–4) | 3 (1–4) | ≤0.01 |
IRS ≤ 8 Final Specimen n = 51 | IRS > 8 Final Specimen n = 12 | ||
Regression grade according to Sinn (median, range) | 4 (0–4) | 1 (0–2) | ≤0.01 |
Difference between IRS Core Biopsy and IRS Final Specimen < 6 n = 39 | Difference between IRS Core Biopsy and IRS Final Specimen ≥ 6 n = 24 | ||
Regression grade according to Sinn (median, range) | 1 (0–4) | 4 (2–4) | ≤0.01 |
Odds Ratio | 95% CI | p | |
---|---|---|---|
Age | 0.01 | −0.02–0.02 | 0.94 |
cT-stage | −0.35 | −0.66–−0.04 | 0.03 |
cN-stage | 0.04 | −0.25–0.32 | 0.78 |
G | 0.36 | −0.28–0.99 | 0.26 |
Ki67 | 0.01 | −0.01–0.02 | 0.85 |
IRS > 8 core biopsy | 0.41 | −0.15–0.96 | 0.15 |
IRS > 8 final specimen | −0.51 | −1.28–0.26 | 0.19 |
Difference between IRS core biopsy and IRS final specimen ≥ 6 | 1.75 | 1.16–2.34 | ≤0.01 |
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Radosa, J.C.; Kasoha, M.; Doerk, M.; Cullmann, A.; Kaya, A.C.; Linxweiler, M.; Radosa, M.P.; Takacs, Z.; Tirincsi, A.; Lang, S.; et al. The 3q Oncogene SEC62 Predicts Response to Neoadjuvant Chemotherapy and Regulates Tumor Cell Migration in Triple Negative Breast Cancer. Int. J. Mol. Sci. 2023, 24, 9576. https://doi.org/10.3390/ijms24119576
Radosa JC, Kasoha M, Doerk M, Cullmann A, Kaya AC, Linxweiler M, Radosa MP, Takacs Z, Tirincsi A, Lang S, et al. The 3q Oncogene SEC62 Predicts Response to Neoadjuvant Chemotherapy and Regulates Tumor Cell Migration in Triple Negative Breast Cancer. International Journal of Molecular Sciences. 2023; 24(11):9576. https://doi.org/10.3390/ijms24119576
Chicago/Turabian StyleRadosa, Julia C., Mariz Kasoha, Merle Doerk, Annika Cullmann, Askin C. Kaya, Maximilian Linxweiler, Marc P. Radosa, Zoltan Takacs, Andrea Tirincsi, Sven Lang, and et al. 2023. "The 3q Oncogene SEC62 Predicts Response to Neoadjuvant Chemotherapy and Regulates Tumor Cell Migration in Triple Negative Breast Cancer" International Journal of Molecular Sciences 24, no. 11: 9576. https://doi.org/10.3390/ijms24119576