Original ArticlesXPC deficiency leads to centrosome amplification by inhibiting BRCA1 expression upon cisplatin-mediated DNA damage in human bladder cancer
Introduction
Chromosome instability is widely accepted as a hallmark of human cancer. The centrosome, a major microtubule-organizing center of mammalian cells, plays fundamental roles in organizing both the interphase cytoskeleton and the mitotic spindle. The presence of more than two centrosomes (centrosome amplification, CA) leads to the formation of defective mitotic spindles and, consequentially, chromosome segregation errors. Due to a strong association between CA and aneuploidy, centrosome amplification is believed to be a major cause of chromosome instability in human cancer cells. Studies have shown the frequent occurrence of centrosome amplification in many types of cancers, including breast, lung, head and neck, prostate, colon, brain, liver and bladder cancer [1]. Moreover, studies on bladder cancer indicated that higher degrees of centrosome amplification were strongly associated with higher tumor stage and tumor recurrence [2,3].
Cellular genomic DNA normally sustains continuous damage requiring repair [4]. It is known that a broad range of DNA-damaging agents, including cytostatic drugs and ionizing radiation, can induce centrosome amplification in both malignant and non-transformed cell lines [[5], [6], [7], [8]]. These different types of DNA damage lead to a consequent DNA damage response (DDR), which orchestrates the repair of the damage, halts cell cycle progression and upregulates the expression of genes involved in the repair process. With DNA damage checkpoint, this cell cycle delay is maintained until the DNA damage is repaired or until the cell is driven out of the cell cycle into apoptosis or senescence [9]. It is well established that the following DDR proteins are present at centrosomes, at least during part of the cell cycle: ATR, ATM, CHK1, CHK2, BRCA1, BRCA2 and PARPs [10].
The nucleotide excision repair (NER) pathway is the major DNA repair pathway for repairing the bulky DNA damage generated by most environmental factors, such as UV radiation, chemicals, and therapeutic drugs. A functional NER pathway is essential for maintaining genetic integrity and for preventing carcinogenesis [11]. Xeroderma pigmentosum group C (XPC) is a DNA damage recognition protein that plays an important role in the NER process. The XPC protein might also play important roles in other DNA damage responses, including cell cycle checkpoint regulation and apoptosis [12]. XPC defects result in the frequent occurrence of mutations in genomic DNA upon DNA damage [13]. XPC defects also have been found in many types of cancer, including lung and skin cancer [14,15]. However, the relationship between XPC defects and chromosomal aberrations remains unclear.
Human bladder cancer is a heterogeneous neoplasm that presents as a superficial tumor in 80% of cases or as a muscle invasive tumor. Molecular biological research revealed that urothelial carcinoma contains multiple genetic and epigenetic abnormalities, including chromosomal abnormalities, oncogene activation, tumor suppressor gene inactivation and tumor microenvironment alterations [16]. We previously reported that attenuated XPC expression was frequent in bladder cancer tissue and was associated with carcinogenesis and progression of bladder cancer [17,18]. Here, we found that attenuated XPC expression was significantly related to centrosome amplification in bladder cancer tissue in this study. This interesting phenomenon led us to investigate how XPC deficiency affects centrosome amplification.
Section snippets
Plasmids, lentivirus and cell lines
For plasmid construction, the cDNA of XPC (Origene) was inserted between the SalI and NotI sites of the pLVX-IRES-Puro vector (Clontech). The Quikchange method (QuikChange II Site-Directed Mutagenesis Kit, Agilent) was used to delete coding region of Centrin2 binding motif of XPC (aa847-863). The promoter region of POU1F1 was inserted between the XhoI and EcoRI sites of the pGL3-basic vector (Promega).
The pLVX-IRES-Puro vector based plasmids were subjected to produce lentivirus particle by
Centrosome amplification in human urothelial carcinoma samples correlates negatively with XPC expression
Based on our previous work, we hypothesized that low XPC expression might contribute to centrosome amplification. To test this hypothesis, centrosome amplification was determined by immunofluorescence staining of γ-tubulin in two panels of bladder cancer tissue selected according to their XPC expression levels. Among the 28 cases selected, 13 cases had high XPC expression, and 15 cases had low XPC expression (Fig. 1A, Upper panel). Centrosome amplification (CA) was detected in all cases (Fig. 1
Discussion
Accumulating evidences indicate that centrosome amplification is a common occurrence and is correlated with malignancy degree and poor prognosis in human bladder cancer [2,3,25]. In our current study, attenuated XPC expression was firstly found to be strongly associated with centrosome amplification in bladder urothelial cells of XPC knockout mice and human bladder cancer. A significant increase in centrosome amplification was observed in XPC-silenced cells upon cisplatin treatment. XPC
Conflicts of interest
The authors declare no potential conflicts of interest.
Acknowledgments
We thank Dr. Ying Wan for his technical support on the Immunofluorescence microscopy studies.
This work was supported by the National Nature Science Foundation of China (Grant number, 30972979, 81372726, 81272822).
More information is available in supplementary files.
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Xeroderma Pigmentosum: A Model for Human Premature Aging
2021, Journal of Investigative DermatologyCitation Excerpt :For example, XPC protein has a role in transcriptional regulation in the cell (Bidon et al., 2018; Le May et al., 2010; Ray et al., 2009; Zebian et al., 2019; Zhu et al., 2009). XPC has a role in maintaining centrosome homeostasis (Wang et al., 2019). XPC deficiency inhibits BRCA1 expression by upregulating its transcriptional repressor, Pit-1.
The Centrosome Linker and Its Role in Cancer and Genetic Disorders
2020, Trends in Molecular MedicineCitation Excerpt :Cancer cells can survive and overcome these surveillance pathways by inhibiting or deleting the tumor-suppressor p53. Although inactivation of p53 seems to dominate the mutational landscape associated with an abnormal number of centrioles, p53 wild-type tumors are also frequently aneuploid, and it is likely that there are other mechanisms that allow the survival of cells with centrosome amplifications [80,81]. Increased Polo-like kinase 4 (PLK4) expression (Box 1) can lead to the acquisition of extra centrosomes via a mechanism involving the centrosomal adaptor proteins CEP152 and CEP192, as well as phosphorylation of STIL, leading to assembly of the SAS-6 cartwheel (Figure 2B) [82–85].