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Author Spotlight: Exploring the Role of FAM83A in Cervical Cancer
Published on: February 9, 2024
Identification and Expression Analysis of CD73 Inhibitors in Cervical Cancer
Jamshed Iqbal1, Ayesha Basharat1, Sehrish Bano1
1Centre for Advanced Drug Research, COMSATS University Islamabad, Abbottabad Campus, Abbottabad-22060, Pakistan.
Aims:
The present study was conducted to examine the inhibitory effects of synthesized sulfonylhydrazones on the expression of CD73 (ecto-5'-NT).
Background:
CD73 (ecto-5'-NT) represents the most significant class of ecto-nucleotidases, which are mainly responsible for the dephosphorylation of adenosine monophosphate to adenosine. Inhibition of CD73 played an important role in the treatment of cancer, autoimmune disorders, precancerous syndromes, and some other diseases associated with CD73 activity.
Objective:
Keeping in view the significance of CD73 inhibitor in the treatment of cervical cancer, a series of sulfonylhydrazones (3a-3i) derivatives synthesized from 3-formylchromones were evaluated.
Methods:
All sulfonylhydrazones (3a-3i) were evaluated for their inhibitory activity towards CD73 (ecto-5'-NT) by the malachite green assay and their cytotoxic effect was investigated on the HeLa cell line using MTT assay. Secondly, the most potent compound was selected for cell apoptosis, immunofluorescence staining, and cell cycle analysis. After that, CD73 mRNA and protein expression were analyzed by real-time PCR and Western blot.
Results:
Among all compounds, 3h, 3e, 3b, and 3c were found to be the most active against rat-ecto- 5'-NT (CD73) enzyme with IC50 (μM) values of 0.70 ± 0.06 μM, 0.87 ± 0.05 μM, 0.39 ± 0.02 μM, and 0.33 ± 0.03 μM, respectively. These derivatives were further evaluated for their cytotoxic potential against cancer cell line (HeLa). Compounds 3h and 3c showed the cytotoxicity at IC50 value of 30.20 ± 3.11 μM and 86.02 ± 7.11 μM, respectively. Furthermore, compound 3h was selected for cell apoptosis, immunofluorescence staining, and cell cycle analysis, which showed a promising apoptotic effect in HeLa cells. Additionally, compound 3h was further investigated for its effect on the expression of CD73 using qRT-PCR and western blot.
Conclusion:
Among all synthesized compounds (3a-3i), Compound 3h (E)-N'-((6-ethyl-4-oxo-4Hchromen- 3-yl) methylene)-4-methylbenzenesulfonohydrazide was identified as the most potent compound. Additional expression studies conducted on the HeLa cell line proved that this compound successfully decreased the expression level of CD73 and thus, inhibited the growth and proliferation of cancer cells.
Insights
Synthesized sulfonylhydrazones were evaluated for their potential to inhibit CD73 (ecto-5'-NT). Compound 3h demonstrated significant CD73 inhibition and reduced cancer cell growth by decreasing CD73 expression.
Area of Science:
- Biochemistry
- Medicinal Chemistry
- Oncology
Background:
- CD73 (ecto-5 -NT) is a key enzyme in adenosine metabolism.
- CD73 inhibition is a promising therapeutic strategy for cancer and autoimmune diseases.
Purpose of the Study:
- To synthesize and evaluate sulfonylhydrazone derivatives as CD73 inhibitors.
- To investigate the anti-cancer effects of these compounds on cervical cancer cells.
Main Methods:
- Synthesis of sulfonylhydrazone derivatives (3a-3i).
- Enzyme inhibition assay (malachite green) and cytotoxicity assay (MTT) on HeLa cells.
- Apoptosis, cell cycle, immunofluorescence, qRT-PCR, and Western blot analyses for the most potent compound.
Main Results:
- Compounds 3h, 3e, 3b, and 3c showed potent CD73 inhibitory activity with IC50 values as low as 0.33 μM.
- Compound 3h exhibited significant cytotoxicity against HeLa cells (IC50 = 30.20 μM) and induced apoptosis.
- Compound 3h effectively reduced CD73 mRNA and protein expression in HeLa cells.
Conclusions:
- Compound 3h, (E)-N'-((6-ethyl-4-oxo-4Hchromen-3-yl) methylene)-4-methylbenzenesulfonohydrazide, is a potent CD73 inhibitor.
- Compound 3h suppresses cancer cell growth by downregulating CD73 expression, indicating therapeutic potential.
