Ahmad Anas, Prakash Ravi, Khan Mohd Shahnawaz, Altwaijry Nojood, Asghar Muhammad Nadeem, Raza Syed Shadab, Khan Rehan
Chemical Biology Unit, Institute of Nano Science and Technology, Knowledge City, Sector-81, Mohali 140306, Punjab, India.
Laboratory for Stem Cell & Restorative Neurology, Department of Biotechnology, Era's Lucknow Medical College Hospital, Sarfarazganj, Lucknow 226003, Uttar Pradesh, India.
ACS Omega. 2022 Apr 4;7(15):13095-13101. doi: 10.1021/acsomega.2c00527. eCollection 2022 Apr 19.
Colorectal cancer (CRC) is one of the major causes of cancer-linked mortality worldwide. Selective therapeutic approaches toward cancer are the need of the hour to combat cancer. Synthetic lethality is a pragmatic targeted cancer therapy in which cancer cell-specific vulnerabilities such as genetic defects/somatic mutations are exploited for selective cancer therapy by targeting genetic interactors (synthetic lethal interactors) of such mutation/defects present in cancer cells. In this study, we investigated the synthetic lethal interaction between checkpoint kinase 2 () and peroxiredoxin-2 () in CRC cells to precisely target CRC cells having defects. We have performed siRNA-mediated silencing and -carbamoyl alanine (NCA)-mediated inhibition of PRDX2 in -null HCT116 cells to confirm the synthetic lethal (SL) interaction between PRDX2 and as the cell population reduced significantly after silencing/inhibition of PRDX2. Additionally, treatment with NCA resulted in an increased level of total ROS in both cell types (HCT116 and -null HCT116 cells), which further confirms that inhibition of PRDX2 results in an increased ROS level, which are mainly responsible for DNA double-strand breaks (DSBs). ROS-induced DNA DSBs get repaired in HCT116 cells, in which is in the normal functional state, but these DNA DSBs persist in -null HCT116 cells as confirmed by the immunofluorescence analysis of 53BP1 and γ-HAX. Finally, -null HCT116 cells undergo apoptosis due to persistent DNA damage as confirmed by immunofluorescence analysis of cleaved caspase-3. The findings of this study suggest that PRDX2 has a SL interaction with , and this interaction can be exploited for the targeted cancer therapy using NCA as a drug inhibitor of PRDX2 for the therapy of colorectal cancer having defects. Further studies are warranted to confirm the interaction in the preclinical model.
结直肠癌（CRC）是全球癌症相关死亡的主要原因之一。当下对抗癌症需要有针对性的癌症治疗方法。合成致死是一种实用的靶向癌症治疗方法，其中利用癌细胞特异性的脆弱性，如基因缺陷/体细胞突变，通过靶向癌细胞中此类突变/缺陷的基因相互作用因子（合成致死相互作用因子）来进行选择性癌症治疗。在本研究中，我们调查了结直肠癌细胞中检查点激酶2（ ）与过氧化物酶体增殖物激活受体γ辅激活因子2（ ）之间的合成致死相互作用，以精确靶向具有 缺陷的结直肠癌细胞。我们在 缺失的HCT116细胞中进行了小干扰RNA（siRNA）介导的 沉默以及N - 氨甲酰丙氨酸（NCA）介导的PRDX2抑制，以确认PRDX2与 之间的合成致死（SL）相互作用，因为在沉默/抑制PRDX2后细胞数量显著减少。此外，用NCA处理导致两种细胞类型（HCT116和 缺失的HCT116细胞）中的总活性氧（ROS）水平升高，这进一步证实了PRDX2的抑制导致ROS水平升高，而ROS主要负责DNA双链断裂（DSB）。ROS诱导的DNA DSB在 处于正常功能状态的HCT116细胞中得到修复，但如通过53BP1和γ - HAX的免疫荧光分析所证实的，这些DNA DSB在 缺失的HCT116细胞中持续存在。最后，如通过裂解的半胱天冬酶 - 3的免疫荧光分析所证实 的， 缺失的HCT116细胞由于持续的DNA损伤而发生凋亡。本研究结果表明，PRDX2与 存在SL相互作用，并且这种相互作用可被用于靶向癌症治疗，使用NCA作为PRDX2的药物抑制剂来治疗具有 缺陷的结直肠癌。需要进一步的研究来在临床前模型中确认这种相互作用。
