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Objective:To investigate the expression of long non-coding RNA (lncRNA) ALMS1-IT1 in colorectal cancer tissues and the molecular mechanism of its effect on the proliferation and migration of colorectal cancer HT-29 cells in vitro.Methods:The cancer tissue specimens and paracancerous tissue (>5 cm from the edge of the tumor) specimens were collected from 40 colorectal cancer patients who were diagnosed by pathological examination after surgical resection in Hubei 672 Orthopedic Hospital of Integrated Traditional Chinese and Western Medicine from July 2018 to November 2020. Real-time quantitative polymerase chain reaction (qRT-PCR) was used to detect the expression level of ALMS1-IT1 in colorectal cancer tissues and paracancerous tissues, when the relative expression of ALMS1-IT1 was higher than or equal to its median relative expression, ALMS1-IT1 was highly expressed, and the correlation of ALMS1-IT1 expression with the clinicopathological characteristics of patients was analyzed. HT-29 cells were infected with the empty lentivirus and the lentivirus carrying the ALMS1-IT1 silence sequence, and named control group and si-ALMS1-IT1 group. qRT-PCR was used to detect the expression of ALMS1-IT1 in the two groups of HT-29 cells. CCK-8 method and Transwell experiment were used to detect the proliferation and migration ability of the two groups of HT-29 cells. The starBase v2.0 online database was used to predict ALMS1-IT1 interacting molecules, and qRT-PCR and Western blot were used to detect the expression of these molecules.Results:The relative expression of ALMS1-IT1 in colorectal cancer tissues was higher than that in paracancerous tissues (4.54±0.61 vs. 1.19±0.31, t = 34.89, P < 0.01). The median relative expression of ALMS1-IT1 in cancer tissues of 40 patients was 2.93, and the high expression rate of ALMS1-IT1 was 50.0% (20/40). The high expression rate of ALMS1-IT1 in cancer tissues of TNM stage Ⅲ patients was higher than that in TNM stage Ⅰ-Ⅱ patients, the high expression rate of ALMS1-IT1 in poorly-differentiated patients was higher than that in well- and moderately-differentiated patients, and the high expression rate of ALMS1-IT1 in patients with lymph node metastasis was higher than that in patients without lymph node metastasis (all P < 0.01). The cell proliferation capacity (absorbance value) of HT-29 cells in the si-ALMS1-IT1 group after cultured for 2, 3, 4, and 5 days was lower than that in the control group (all P < 0.05). The number of cell migration at 24 h in HT-29 cells of the si-ALMS1-IT1 group was less than that of the control group (45±7 vs. 112±18, t = 3.45, P < 0.05). Using starBase v2.0 online database to predict that the target gene of ALMS1-IT1 may be miRNA-889-3p (miR-889-3p), and the target gene of miR-889-3p may be ATAD2. Compared with the control group, the relative expression of miR-889-3p in HT-29 cells of the si-ALMS1-IT1 group increased (4.24±0.46 vs. 1.01±0.11, t = 6.81, P < 0.01). Compared with the control group, ATAD2 mRNA ( P < 0.01) and protein expression levels in the si-ALMS1-IT1 group were reduced. Conclusions:ALMS1-IT1 is highly expressed in colorectal cancer tissues, and the ALMS1-IT1 expression is related to the TNM stage, degree of tumor differentiation and lymph node metastasis of patients. Down-regulation of ALMS1-IT1 in vitro may inhibit the proliferation and migration of colorectal cancer HT-29 cells by regulating the miR-889-3p-ATAD2 axis. ALMS1-IT1 may be a therapeutic target for colorectal cancer.
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Objective To investigate the expressions of ATPase family AAA domain-containing protein 2 (ATAD2) and β-catenin,and to analyze their correlations with clinicopathological features and prognostic significance in patients with hepatocellular carcinoma (HCC).Methods The HCC tissues of 40 patients were tested by real-time PCR to study the expressions of ATAD2 and β-catenin.Real-time PCR and Western blot were performed to detect the proteins and mRNA levels of ATAD2,APC,β-catenin and wnt signaling pathway downstream.The HCC tissues of 80 patients and 20 peritumoral tissues were tested by immunohistochemistry (IHC).The cumulative survival-rate was analyzed by the Kaplan-Meier method,and univariate and multivariate were analyzed by the Cox proportional hazards model.Results ISH:The positive rates of ATAD2 and β-catenin were 65.0％ and 55.0％,respectively.These rates were significantly higher than those in the peritumoral tissues (30.0％ and 25.0％,respectively).The ATAD2 expression was related to tumor size (P ＜ 0.05),metastasis (P ＜ 0.05),serum AFP level (P ＜ 0.05) and TNM stag ing (P ＜ 0.05).The β-catenin expression was only significantly related to metastasis (P ＜ 0.05).Correlation analysis showed that the ATAD2 expression was positively related to the β-catenin expression (Pearson =0.578,P ＜ 0.01,R2 =0.3607,Spearman =0.495).This positive relationship was also found in the remaining 4 cell lines except the SK-hep1.Depleting ATAD2 up-regulated APC and down-regulated β-catenin protein and mRNA expression.Univariate and multivariate analyses showed that ATAD2 and β-catenin expressions,tumor size,metastasis,serum AFP,and TNM staging were poor prognostic factors for HCC,and ATAD2 and β-catenin expressions,metastasis,serum.AFP were independent prognostic factors.Patients whose ATAD2 and β-catenin were both positive had worse survival than those with only one positive expression or both negative expressions (P ＜ 0.05).Depleting ATAD2 down-regulated survivin,cyclinD1,c-myc,MMP7,Vimentin in wnt signaling pathway and EMT related proteins.Conclusions ATAD2 and β-catenin expressions were positively related in patients with HCC.Abnormal expressions between ATAD2 and β-catenin might participate in the wnt signaling pathway.
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Objective@#To examine the expression of ATAD2 in different liver lesions and its clinical significance.@*Methods@#ATAD2 expression in 60 hepatocellular carcinoma (HCC) surgical specimens (49 of which have concurrent liver cirrhosis), 43 HCC biopsy specimens, 2 high-grade liver dysplastic nodule specimens, 3 low-grade liver dysplastic nodule specimens, 50 liver cirrhosis tissue samples, and 20 normal liver tissue samples were measured using immunohistochemistry. The F-test, q-test, t-test, and chi-square test were used for statistical analysis of data.@*Results@#ATAD2 was expressed in 56 HCC surgical specimens (93.33%), 35 HCC biopsy specimens (81.40%), and 2 high-grade liver dysplastic nodule specimens (2/2), but not in the low-grade liver dysplastic nodule, liver cirrhosis tissue, and normal liver tissue samples. The mean expression of ATAD2 was significantly higher in HCC tissues than in high-grade and low-grade liver dysplastic nodule tissues, liver cirrhosis tissue, and normal liver tissue (F = 22.96, q = 3.138, 3.972, 12.272, and 9.101, respectively, all P < 0.01). There were no significant differences in the mean expression and positive expression rate of ATAD2 between HCC surgical and biopsy specimens (t = 1.40, P > 0.05; χ² = 3.47, P >0.05). Of the 35 HCC biopsy specimens that expressed ATAD2, the mean ATAD2 expression was ≥1% in 35 specimens (100%), ≥3% in 27 specimens (77.14%), and ≥5 % in 23 specimens (65.71%). In addition, among the pathological grade I-II HCC biopsy specimens, the mean ATAD2 expression was ≥1% in 28 specimens (100%), ≥3% in 22 specimens (62.86%), and ≥5% in 19 specimens (54.29%). Moreover, ATAD2 expression in HCC was associated with serum alpha-fetoprotein level, presence of hepatitis B virus surface antigen (HBsAg), and presence of concurrent liver cirrhosis (t = 2.09, 2.30, and 2.18, respectively, all P < 0.05).@*Conclusion@#ATAD2 may play an important role in HCC tumorigenesis, and may be involved in malignant transformation of cells. ATAD2 expression can be a valuable marker for differentiating the nature of lesions in liver biopsy tissues during clinical practice.
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Objective To analyze the effect of ATAD2 on glioma cell lines.Methods ATAD2 level in U87MG, U251 and normal human astrocytes was detected by RT-PCR and Western blot.U87MG and U251 cells were divided into four groups: control, mock (lipofection control), ATAD2 siRNA (transfected with ATAD2 siRNA) and ATAD2 overexpression (transfected with ATAD2).Cell proliferation, migration and invasion were respectively measured by MTT and Transwell assay.The level of E-cadherin, N-cadherin and Vimentin was measured by Western blot.Results ATAD2 was highly expressed in U87MG and U251 cells.Compared with control, cell proliferation, invasion, migration, N-cadherin and Vimentin expression were decreased by ATAD2 siRNA, while they were promoted by ATAD2 overexpression (P<0.05).E-cadherin expression was upregulated by ATAD2 siRNA and it was inhibited by ATAD2 overexpression (P<0.05).Conclusions ATAD2 participants in human glioma cells metastasis via epithelial-mesenchymal transition (EMT).
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Objective To investigate the correlation and clinicopathological features of ATAD2 and E-cadherin expression in human hepatocellular carcinoma (HCC),and role of ATAD2 in HCC cells invasion and metastases.Methods The expressions of ATAD2 and E-cadherin in 90 HCC specimens and 16 specimens collected from peritumoral tissue were detected by immunohistochemistry (IHC) test.The correlations between ATAD2 and E-cadherin expression and clinicopathological features were analyzed.The lentiviral vector of sh-ATAD2 RNA was used to interfere with ATAD2 expression.Western blot was performed to evaluate the changes of E-cadherin and Vimentin expression in HCC cells after ATAD2 expression interference.The role of ATAD2 in HCC cells invasion and metastasis was assessed by Transwell analysis.Results ATAD2 protein expression was obviously upregulated in HCC tissue compared to peritumoral tissue (56.7％ vs.31.3％,P ＜ 0.05).Meanwhile,E-cadherin expression was significantly downregulated in HCC tissue (43.3％ vs.68.7％,P ＜0.05).ATAD2 and E-cadherin expressions were both correlated with HCC metastasis (P ＜ 0.05).Correlation analysis demonstrated that the ATAD2 expression was negatively correlated with E-cadherin (r =-0.263,P ＜ 0.05).After interfering ATAD2 expression,cell migration and invasion of HCCLM3 cells were suppressed (migration,42.5 ± 2.6 vs.78.1 ± 3.8,P ＜ 0.05;invasion,33.0 ± 4.7 vs.72.7 ± 5.6,P ＜ 0.05).Meanwhile,the E-cadherin expression was remarkably upregulated (P ＜ 0.05).However,the Vimentin expression downregulated (P ＜ 0.05).Conclusions ATAD2 expression was upregulated in HCC,and silencing ATAD2 could upregulate E-cadherin expression and downregulate Vimentin expression,and inhibit the invasion and metastasis of HCC cells.Modulating E-cadherin expression may be the potential mechanism of ATAD2 in HCC invasion and metastasis.
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PURPOSE: Cancer cells frequently express genes that are specifically or preferentially expressed in male germ cells under normal conditions. The ATPase family AAA domain-containing 2 (ATAD2) is one such and works as an important cofactor for MYC-dependent transcription. In hepatocellular carcinoma (HCC), ATAD2 has been identified as a candidate driver gene located within the amplified 8q24 locus. However, the prognostic significance of ATAD2 protein expression in HCC remains uncertain. MATERIALS AND METHODS: We investigated ATAD2 protein expression by immunohistochemistry in tumor tissue from 182 HCC patients who underwent curative resection. Associations of ATAD2 expression with clinicopathologic variables or prognosis of HCC patients were analyzed. RESULTS: ATAD2 expression was observed in 119 (65.4%) of the 182 HCCs and tended to be independent predictor of early recurrence (p=0.059). ATAD2 expression showed an unfavorable influence on recurrence-free survival (RFS) (p < 0.001). Subgroup analysis among patients with tumor size < or = 5.0 cm (n=109), patients at Barcelona Clinic Liver Cancer stage 0 or A (n=92), and patients with alpha-fetoprotein < or = 20 ng/mL (n=61), the ATAD2-positive groups unfavorably influenced RFS (p=0.008, p=0.009, and p=0.013, respectively). In addition, ATAD2 expression was an independent predictor of shorter RFS (p=0.002). ATAD2 expression showed an unfavorable influence on disease-specific survival (p=0.001), but was not an independent predictor of shorter disease-specific survival (p=0.109). CONCLUSION: ATAD2 protein expression may be a potential predictor of RFS in HCC patients after curative resection and ATAD2 may have prognostic value in patients with early stage HCC or normal serum alpha-fetoprotein level.
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Humans , Male , Adenosine Triphosphatases , alpha-Fetoproteins , Carcinoma, Hepatocellular , Germ Cells , Immunohistochemistry , Liver Neoplasms , Prognosis , RecurrenceABSTRACT
AT Pase family, AAA domain containing 2 (ATAD2) is a newly uncovered oncogene playing an important role in occurrence and development of prostate cancer, breast cancer and other human malignancies. It is a coactivator of a variety of transcription factors, and it can also modulate genes through epigenetic modifications, resulting in overexpression of its downstream oncogenes, and in turn changing the phenotypes of tumor cells; what's more, the expression of ATAD2 is associated with histological grade of cancer, metastasis, disease recurrence and the survival, indicating that it can be used as a molecular marker for prognosis of cancer and has remarkable value in clinical application. Furthermore, ATAD2 contains bromodomain (BRD) and also functions as cellular activity-associated ATPase, which means that ATAD2 may be a potential target for drug therapy. This review summarizes the recent findings and discusses the prospect of this field.