The cell cycle is a rich field for research, especially, the DNA damage. DNA damage, which happened naturally or as a result of environmental influences causes change in the chemical structure of DNA. The extent of DNA damage has a significant impact on the fate of the cell in later stages. In this paper, we introduced an Unsupervised Machine learning Model for DNA Damage Diagnosis and Analysis. Mainly, we employed K-means clustering unsupervised machine learning algorithms. Unsupervised algorithms commonly draw conclusions from datasets by solely utilizing input vectors, disregarding any known or labeled outcomes. The model provided deep insight about DNA damage and exposes the protein levels for proteins when work together in sub-network model to deal with DNA damage occurrence, the unsupervised artificial model explained the sub-network biological model activities in regard to the changing in their concentrations in several clusters, they have been grouped in such as (0 – no damage, 1 – low, 2 – medium, 3 – high, and 4 – excess) DNA damage clusters. The results provided a rational and persuasive explanation for numerous important phenomena, including the oscillation of the protein p53, in a clear and understandable manner. Which is encouraging since it demonstrates that the K-means clustering approach can be easily applied to many similar biological systems, which aids in better understanding the key dynamics of these systems.