Small Molecule Inhibitors Targeting Key Proteins in the DNA Damage Response for Cancer Therapy.

DNA damage response (DDR) is a complicated interactional pathway. Defects that occur in subordinate pathways of the DDR pathway can lead to genomic instability and cancer susceptibility. Abnormal expression of some proteins in DDR, especially in the DNA repair pathway, are associated with the subsistence and resistance of cancer cells. Therefore, the development of small molecule inhibitors targeting the chief proteins in the DDR pathway is an effective strategy for cancer therapy. In this review, we summarize the development of small molecule inhibitors targeting chief proteins in the DDR pathway, particularly focusing on their implications for cancer therapy. We present the action mode of DDR molecule inhibitors in preclinical studies and clinical cancer therapy, including monotherapy and combination therapy with chemotherapeutic drugs or checkpoint suppression therapy.

MicroRNA-140 impedes DNA repair by targeting FEN1 and enhances chemotherapeutic response in breast cancer.

An increased DNA repair capacity is associated with drug resistance and limits the efficacy of chemotherapy in breast cancers. Flap endonuclease 1 (FEN1) participates in various DNA repair pathways and contributes to cancer progression and drug resistance in chemotherapy. Inhibition of FEN1 serves as a potent strategy for cancer therapy. Here, we demonstrate that microRNA-140 (miR-140) inhibits FEN1 expression via directly binding to its 3' untranslated region, leading to impaired DNA repair and repressed breast cancer progression. Overexpression of miR-140 sensitizes breast cancer cells to chemotherapeutic agents and overcomes drug resistance in breast cancer. Notably, ectopic expression of FEN1 abates the effects of miR-140 on DNA damage and the chemotherapy response in breast cancer cells. Furthermore, the transcription factor/repressor Ying Yang 1 (YY1) directly binds to the miR-140 promoter and activates miR-140 expression, which is attenuated in doxorubicin resistance. Our results demonstrate that miR-140 acts as a tumor suppressor in breast cancer by inhibiting FEN1 to repress DNA damage repair and reveal miR-140 to be a new anti-tumorigenesis factor for adjunctive breast cancer therapy. This novel mechanism will enhance the treatment effect of chemotherapy in breast cancer.

Enhanced Activity of Variant DNA Polymerase ß (D160G) Contributes to Cisplatin Therapy by Impeding the Efficiency of NER.

Cisplatin, commonly used in a variety of cancer treatments, induces apoptosis in cancer cells by causing lethal DNA damage. Several DNA repair pathways participate in regulation of cisplatin treatment, leading to cisplatin sensitivity or resistance in cancer cells. DNA polymerase ß (pol ß), a key protein involved in base excision repair, confers a response to cisplatin therapy that is dependent on polymerase activity. Pol ß D160G mutation with enhanced polymerase activity, previously identified in clear cell renal cell carcinoma, enhances the sensitivity of human cancer cells and mouse xenografts to cisplatin by limiting the efficiency of nucleotide excision repair (NER). Notably, the D160G mutation impedes the recruitment of XPA to cisplatin-induced sites of DNA damage, leading to unrepaired damage and further inducing cell death. Molecular architecture analysis indicated that the D160G mutation alters protein-DNA interactions and the surface electrostatic properties of the DNA-binding regions, resulting in greater DNA affinity and polymerase activity compared with wild-type pol ß. Collectively, these results indicate that enhancing pol ß activity impedes the efficiency of NER and provide a promising adjuvant therapeutic strategy for cisplatin chemotherapy. IMPLICATIONS: Our studies demonstrate that polß D160G mutation with enhanced polymerase activity impedes NER efficiency during the repair of cisplatin-induced DNA damage, leading to increased cisplatin sensitivity in cancer cells.

Obesity-induced overexpression of miRNA-24 regulates cholesterol uptake and lipid metabolism by targeting SR-B1.

Scavenger Receptor B1 (SR-B1) is an 82 kDa integral membrane glycoprotein that mediates selective uptake of high-density lipoprotein cholesteryl ester (CE) in vitro and in vivo. Previously, we defined several kinds of regulatory mechanisms of SR-B1 expression and function. Here, we have dissected the function of a novel miR-24 on SR-B1 expression, HDL uptake and lipid metabolism. We showed that miR-24 was upregulated in HepG2 cells cultured in the mimicked hyperlipidemic condition and in the livers of dietary induced and genetic obesity mice. Overexpression of miR-24 inhibited SR-B1 expression by directly targeting SR-B1 3' UTR and repressed HDL uptake and steroidogenesis in steroidogenic cells. HepG2 cells with miR-24 showed attenuation of TG levels and lipid accumulation. Moreover, we validated that overexpression of miR-24 downregulated the expression of certain genes involved in lipogenesis, FASN, ACLY and SCD1, and increased the expression of genes of cholesterol synthesis, HMGCR, DHCR24 and SREBP2. Taken together, we demonstrated that obesity induced miR-24 repressed HDL uptake, steroid hormone synthesis and lipid metabolism by targeting SR-B1.

Synergistic antitumor effect of combined paclitaxel with FEN1 inhibitor in cervical cancer cells.

Studies on cervical cancer are urgently required to improve clinical outcomes. As a major anticancer drug for cervical cancer, paclitaxel has been used for many years in clinical therapy but its therapeutic efficacy is limited by common obstacle from cancer cells. The enhanced DNA repair pathways of cancer cells have been proved to survive DNA damage induced by chemotherapeutic drug. Inhibitors of specific DNA repair pathway can sensitize cancer cells to the treatment of chemotherapeutic drugs. In this paper we found that the effect of paclitaxel can be significantly improved when used in combination with FEN1 inhibitor SC13, suggesting a synergistic mechanism between the two compounds. Our studies suggest that FEN1 inhibition could be a novel strategy of tumor-targeting therapy for cervical cancer. Our work also revealed that paclitaxel demonstrates stronger synergistic effect with SC13 than other common used chemical drugs such as doxorubicin, carboplatin or camptothecin on cervical cancer cells.

A comprehensive analysis of plausible genotoxic covariates among workers of a polyvinyl chloride plant exposed to vinyl chloride monomer.

The aim of this study was to assess the frequency of chromosomal aberrations-including chromatid type aberrations (CTAs), chromosomal type aberrations, micronucleus (MN) comet assay, and XRCC1 399 Arg/Gln polymorphism-in peripheral blood lymphocytes of workers occupationally exposed to vinyl chloride monomer (VCM). A total of 52 workers and an equal number of controls were recruited into the study to explore the potential cytogenetic risk of occupational exposure to VCM. Questionnaires were administered to obtain details of habitual cigarette-smoking, alcohol-consumption pattern, and occupation, etc. The exposed subjects and controls were classified into two groups based on age (group I <40 years; group II ≥40 years), and exposed subjects were further classified based on exposure duration (>8 and ≥8 years). CTA, MN, and comet assay frequency were significantly greater in polyvinyl chloride (PVC) factory workers (p < 0.05) with long-duration work. CTA, MN, and comet assay values were found to be increased with age in exposed subjects as well as in controls, with exposed subjects showing a statistically greater degree. An extensively greater MN frequency was observed in smokers exposed to VCM than in the control group (P < 0.05). The mean tail length of exposed subjects was greater compared with controls. The study on XRCC1 399 Arg/gln polymorphism in PVC factory workers showed less significant difference in allele frequency compared with controls. In conclusion, this results of work provides evidence for an apparent genotoxic effect associated with VCM exposure. Our results reinforce the greater sensitivity of cytogenetic assays for biomonitoring of occupationally exposed populations. Statistics indicate that workers exposed to VCM are at carcinogenic risk and should be monitored for long-term adverse effects from their exposure.

Cytogenetic methods for assessing human exposure to toluene in Coimbatore, South India.

The aim of the present study was to identify genetic alterations occurring in rotograving workers chronically exposed to toluene. A total of 60 samples from 30 exposed subjects and 30 age-matched controls, selected based on the toluene level present in their urine, were recruited. Exposed subjects were categorized based on their duration of exposure and smoking habitats. Controls were normal and healthy and categorized based on their smoking habits. Cell cultures were established from blood samples collected from the control and experimental subjects after obtaining informed consent. G-banding and comet assays were used to identify genetic alterations. A higher degree of total chromosome aberration was identified in exposed subjects compared to controls. As expected, controls exhibited minimal number of alterations. The overall CA frequency due to toluene exposure was significantly different from that of the controls for both chromatid and chromosome type aberrations (P\0.05 by ANOVA). . The habit of cigarette smoking among the workers had a synergistic effect on inducing DNA damage. In conclusion, this work shows a clear genotoxic effect associated with toluene exposure, our results also reinforcing the conclusion of higher sensitivity of cytogenetic assays for the biomonitoring of occupationally exposed populations. There is a strong need to educate those who work with potentially hazardous materials about adverse effects and highlight the importance of using protective measures.