Traditional Chinese medicine (TCM), as a significant complementary and alternative approach, has received increasing attention from medical practitioners worldwide and is now regarded as an important source for modern drug discovery owing to its special therapeutic efficiency and few side effects. In this work, a popular TCM, Ganoderma lingzhi (G. lingzhi), was used as a test drug to show how Raman spectroscopy combined with traditional chemical and biological assays could be used to monitor alternations in molecular events in G. lingzhi extract-induced cells with high sensitivity. The observations indicated that the Raman fingerprint of MDA-MB-231 cells after G. lingzhi triterpene treatment was significantly different in contrast to that of the control, and changes in the Raman bands showed that some important molecular events could be discerned, including protein denaturation, DNA fragmentation, and lipid metabolism disorder. Network pharmacology analysis further supplemented the drug's target and signaling pathways. Our work proposes an effective way for screening potential new drug products early from the complex chemical compositions of TCM, as well as for accelerating preclinical medicinal chemistry optimization cycles, playing a significant role in the development of new drugs. [ABSTRACT FROM AUTHOR]