Pactamycin, although putatively touted as a potent
antitumor agent, has never been used as an anticancer
drug due to its high cytotoxicity. In this study, we characterized the effects of two novel biosynthetically engineered analogs of
pactamycin, de-6MSA-7-demethyl-7-deoxypactamycin (TM-025) and 7-demethyl-7-deoxypactamycin (TM-026), in
head and neck squamous cell carcinoma (
HNSCC) cell lines SCC25 and SCC104. Both
TM-025 and
TM-026 exert growth inhibitory effects on
HNSCC cells by inhibiting cell proliferation. Interestingly, unlike their parent compound
pactamycin, the analogs do not inhibit synthesis of nascent
protein in a cell-based assay. Furthermore, they do not induce apoptosis or autophagy in a dose- or a time-dependent manner, but induce mild senescence in the tested cell lines. Cell cycle analysis demonstrated that both analogs significantly induce cell cycle arrest of the
HNSCC cells at S-phase resulting in reduced accumulation of G2/M-phase cells. The
pactamycin analogs induce expression of
cell cycle regulatory proteins including master regulator p53, its downstream target p21Cip1/WAF1, p27kip21, p19,
cyclin E, total and phospho Cdc2 (Tyr15) and Cdc25C. Besides, the analogs mildly reduce
cyclin D1 expression without affecting expression of
cyclin B, Cdk2 and Cdk4. Specific inhibition of p53 by
pifithrin-α reduces the percentage of cells accumulated in S-phase, suggesting contribution of p53 to S-phase increase. Altogether, our results demonstrate that
Pactamycin analogs
TM-025 and
TM-026 induce senescence and inhibit proliferation of
HNSCC cells via accumulation in S-phase through possible contribution of p53. The two PCT analogs can be widely used as research tools for cell cycle inhibition studies in proliferating
cancer cells with specific mechanisms of action.