Abstract:
A. Hit evolutionary hypothesis:
Alphabetic code for cancer management, based on the comprehensive strategic network by considering single cell based analysis/pedigree platform/ Micro-,macro/environmental factors which have significant role through the pre- and post-embryonic periods.
Cancer translational insight relies on the diverse or harmonic behavior at functional and molecular levels. So, It is crucial to unmask the sequential/constructive/cascade events at genomics/tumor levels.
Detecting D1853N polymorphism of Ataxia telangiectasia mutated gene (ATM) gene is the key target in cancer- and non-cancerous individuals, inherited, or as a de novo alteration, located at 11q22q23 and Involved in DNA repair pathway and cell cycle checkpoint.
Material and methods: Includes molecular diagnosis by PCR/sequencing, protein expression (PE) assay at single cell level, and In sillico analysis.
Introduction:
Cell cycle outlines the initiation/progression and therapeutic approaches of neoplasms. An uncontrolled cell proliferation and growth are the key characteristics of neoplasms. Normal checkpoints regulate the machinery of phases through the barriers. So, balancing the oncogenic processes inhibit progression and facilitates the personalized therapy.
Results/discussion:
Three-hit includes D1853N polymorphism, as the first predisposing hit, IVS 35- 63T→A as second hit deriving from the first somatic evolution before differentiation and third-hit (IVS35- 30 A→G) through the tumor development.
Five- hit in breast cancer (BC) includes IVS 36-91 AA>TT, IVS 36-8 T>C, D1853N, IVS 37+47 A>G, IVS 37+60 Del T. IVS 36-8 T>C and D1853N in blood and tumor tissue.Splicing variants occurred at tumor level. Missense D1853N was effective on 2D and 3D structure of ATM protein. PE of ATM also confirmed the functional alterations. Conclusively, five-hit influence the guard of genomic stability, at molecular/cellular/structural levels.
Eight-hit includes D1853N (1st hit), IVS 36-8 T>C as 2nd hit, V1833M as 3rd hit (at pre-differentiation stage), followed by L1888L as the 4th, and somatic variants including IVS 36-46 C>T, L1842L, H1864H, and S1872R, as 5-8th hits. Low PE of ATM was also confirmed, by the diverse expression of cyclin E, CDC25A, P53, and Ki-67.
B. Evolutionary Mosaic phases in cell cycle of BC patients:
Methods: Evolution was traced in interphase to detect the Mosaic Phases (MPs) by Fluorescence In Situ Hybridization (FISH) and PE including immunofluorescence and flow cytometry.
Results: Novel hypothesis reflects the incidence of dual and/or multi-phases, as minor clones in single cells of BC patients. This definition initiated a model, based on the ratio and diverse MPs comprising G1/S, S/G2 and G1/S/G2, and normal phases (G1, S, G2). Significant-harmonic manner was traced between: 1) signal copy numbers, 2)equivalent PE, dual- and triple- co-expression of the cyclins E/B1, D1/E/B1. The ratio between gain/normal signals led to a good prognosis for chromosome 1, but longer survival related to this ratio in chromosome 3.
Key Words: Cell cycle; Mosaic Phases; evolutionary hypothesis; breast cancer; FISH
Conclusions:
A. Hit-hypothesis provides:Reliable platform for early detection/tracing Predisposing/ Prediction/ Prognostic/Preventive (4xP) packages for clinical management.
B. The cell cycle based panel innovates the CDKs inhibitor-based therapy by considering MPs Model in BC and other cancers.
