Abstract
The pro-apoptotic Bax isoform Bax∆2 was originally discovered in cancer patients with a microsatellite guanine deletion (G8 to G7). This deletion leads to an early stop codon; however, when combined with the alternative splicing of exon 2, the reading frame is restored allowing production of a full-length protein (Bax∆2). Unlike the parental Baxα, Bax∆2 triggers apoptosis through a non-mitochondrial pathway and the expression in human tissues was unknown. Here, we analyzed over 1000 tissue microarray samples from 13 different organs using immunohistochemistry. Bax∆2-positive cells were detected in all examined organs at low rates (1–5%) and mainly scattered throughout the connective tissues. Surprisingly, over 70% of normal colon samples scored high for BaxΔ2-positive staining. Only 7% of malignant colon samples scored high, with most high-grade tumors being negative. A similar pattern was observed in most organs examined. We also showed that both Baxα and Bax∆2 can co-exist in the same cells. Genotyping showed that the majority of Bax∆2-positive normal tissues contain no G7 mutation, but an unexpected high rate of G9 was observed. Although the underlying mechanism remains to be explored, the inverse correlation of Bax∆2 expression with tissue malignancy suggests that it may have a clinical implication in cancer development and treatment.
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Acknowledgements
We would like to thank Dr. John Hart from the University of Chicago (Department of Pathology) for his assistance regarding tumor pathology, and Dr. Li Ma from Mumetel LLC for providing his expertise for microsatellite genotyping. We also would like to thank our students Jodi Curtin, Irena Grauzinis and Ana Marija Fonceva, for their contribution. Imaging work was performed at the Northwestern University Center for Advanced Microscopy generously supported by NCI CCSG P30 CA060553 awarded to the Robert H. Lurie Comprehensive Cancer Center.
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This work was supported by the National Institutes of Health R15 CA195526.
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AM, HHZ and JX contributed to experimental design. AM, QY, AD, EB, JL, and AN performed the experiments. AM and SB performed the computational analysis of the tissues. AM, HYZ, QY, HHZ and JX contributed to data analysis and interpretation. AM and JX wrote the manuscript. All authors reviewed the results and approved the final manuscript.
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Dr. Xiang has a patent related to this project but has not received any financial compensation in association with the patent or this work. All authors declare no conflict of interests.
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Mañas, A., Yao, Q., Davis, A. et al. Immunohistochemical detection of the pro-apoptotic Bax∆2 protein in human tissues. Histochem Cell Biol 154, 41–53 (2020). https://doi.org/10.1007/s00418-020-01874-w
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DOI: https://doi.org/10.1007/s00418-020-01874-w