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OBJECTIVE@#To investigate the expression of CDC25A in non- small cell lung cancer (NSCLC) tissues and explore its correlation with the clinicpathological features of the patients and the expressions of let-7a1 and let-7c.@*METHODS@#We collected surgical specimens of pathologically confirmed NSCLC tissues and paired adjacent lung tissues from 44 patients and tissues of benign lung lesions from 9 patients. The expressions of CDC25A protein and mRNA in the tissues were detected by immunohistochemistry and fluorescence quantitative RT-PCR, respectively; the expressions of let-7a1 and let-7c mRNA were detected using tail-adding fluorescence quantitative RT-PCR.@*RESULTS@#The positivity rate of CDC25A protein expression was significantly higher in NSCLC tissues than in the adjacent tissues and benign pulmonary lesions (@*CONCLUSIONS@#The expression level of CDC25A is significantly increased in NSCLC with a negative correlation with Let-7c expression, which identifies CDC25A as a possible downstream target gene of Let-7c.
Subject(s)
Humans , Carcinoma, Non-Small-Cell Lung/genetics , Gene Expression Regulation, Neoplastic , Lung , Lung Neoplasms/genetics , Lymphatic Metastasis , MicroRNAs , RNA, Messenger/genetics , cdc25 PhosphatasesABSTRACT
@#Objective: To explore the genes that may be regulated by cell division cycle 25A (CDC25A) with gene chip technology, and to elucidate and verify that CDC25A has a regulatory effect on the expression of liver cancer related genes. Methods: CDC25A expression in human liver cancer HepG2 cells was silenced by siRNA interference technology and a nude mouse xenograft model of liver cancer was successfully constructed in our previous research. Affymetrix human gene expression profiling microarray was used to further screen differentially expressed genes (DEGs) after silencing CDC25A in liver cancer xenografts, and GO analysis and KEGG analysis were performed. Some of the DEGs were verified by qPCR. Results: The chip screened 188 DEGs in liver cancer xenograft tissues after CDC25A silence, including 78 up-regulated genes and 110 down-regulated genes. These DEGs mainly involved in cell proliferation, apoptosis, protein complex binding, extracellular space, etc., and associated with the changes in pathways such as focal adhesions and extracellular matrix (ECM) receptor interactions. qPCR showed that the expression of HIPK2 mRNA was up-regulated and the mRNA expressions of (microfibrillar-associated protein 5(MFAP5) and cyclin D1 (CCND1) were down-regulated, which were consistent with the results of microarray detection. Conclusion: Using human gene expression profiling chip, the DEGs in liver cancer xenograft tissues in nude mice after silencing CDC25Awere successfully screened, providing effective clues for exploring the effect of CDC25Aon the growth of liver cancer.
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Objective To study the relationship between CDC25A (cell division cycle 25A) expression and the development of gastric adenocarcinoma. hTe effect of artesunate (Art) on CDC25A and gastric cancer cells were also investigated.Methods hTe CDC25A protein expression in gastric adenocarcinoma was detected by lfow cytometry assay. SGC-7901 cells were divided into four groups: control group and 30, 60, 120μmol/L Art groups. Cell apoptosis, cell cycle and CDC25A protein expression in SGC-7901 cells were determined by lfow cytometry atfer the treatment of different concentrations of Art (30, 60, 120μmol/L) for 24h, while the same volume of saline was used in the control.Results CDC25A protein expression level in gastric adenocarcinoma (419.69±21.91) was signiifcantly higher than that in normal gastric tissues (316.11±24.23,P<0.01). hTe cell apoptosis rates of 30, 60, 120μmol/L Art groups (5.48%±0.67%, 12.55%±1.17%, 23.43%±2.18%) were significantly higher than that of control group (0.87%±0.14 %,P<0.05), with an Art dose dependent manner. hTe cell proliferation indices of 30, 60, 120μmol/L Art groups (39.18%±0.53%, 35.71%±0.99%, 31.73%±1.02%) were signiifcantly lower than that of control group (44.12%±2.51%,P<0.01). hTe CDC25A protein expression levels of 30, 60, 120μmol/L Art groups (414.80±4.06, 397.86±3.61, 345.68±7.11) were significantly lower than that of control group (433.99±1.56,P<0.01).ConclusionhTe abnormally increased expression level of CDC25A may be involved in the development of gastric adenocarcinoma. Art can inhibit the growth of SGC-7901 cells by down-regulating the expression of CDC25A protein.
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[ ABSTRACT] AIM:To investigate the effect of silencing cell division cycle 25a ( CDC25a) gene on the prolifera-tion of human hepatoma HepG2 cells.METHODS:CDC25a gene in human hepatoma HepG2 cells was silenced by RNA interference.Real-time PCR was applied to detect the expression of CDC25a, cyclin E and CDK2 at mRNA levels in the HepG2 cells.Western blotting was applied to detect the expression of CDC25a at protein level.In addition, MTT assay, Giemsa staining and flow cytometry were used to measure the proliferation of human hepatoma HepG2 cells.RESULTS:The expression of CDC25a at mRNA and protein levels in RNA silence group was lower than those in negative control group and normal control group (P<0.05).The mRNA expression of cyclin E and CDK2 in silence group was lower than that in negative control group and normal control group (P<0.05).The cell proliferation in silence group was lower than that in negative control group and normal control group ( P<0.05) .The results of flow cytometry revealed that the cells in silence group were blocked in G1 phase.CONCLUSION:Infection of LV-CDC25a-RNAi recombinant to the HepG2 cells effec-tively inhibits the CDC25a gene expression and the proliferation of human hepatoma cells, and arrests the cells in G1 phase, suggesting that CDC25a gene may be a key target for the treatment of liver cancer.
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OBJECTIVE: This study was undertaken to quantitatively detect Cdc25A, Cdc25B and Cdc25C in cervical carcinoma and determine the relationship between the expression of mRNA and protein of cell division cycle (Cdc)25 phosphatase and various clinicopathologic prognostic factors of cervical carcinoma. METHODS: 39 patients diagnosed with cervical carcinoma between February 2000 to March 2005 and 10 patients with benign gynecologic disease were enrolled in this study. A reverse transcriptase-polymerase chain reaction (RT-PCR) and Western blot analysis were used to analyze the expression of Cdc25 phosphatase mRNA and protein in fresh invasive cervical cancer tissue and normal cervix tissue. RESULTS: The mRNA expressions of Cdc25A, Cdc25B and Cdc25C in the cancer tissues were significantly greater than in the control (p=0.02, 0.01, 0.02), respectively. A Western blot analysis yielded same results (p=0.01, 0.02, 0.01). There were also significant relationships between the age and the Cdc25B mRNA expression (p=0.03), between the cell type and the Cdc25C mRNA expression (p=0.04). However, other clinicopathologic prognostic factors including stage, subtype, SCC Ag level, DNA flow cytometry, lymph node metastasis, lymphovascular space invasion and HPV positivity were not statistically significant. CONCLUSION: Our results show that Cdc25A, Cdc25B and Cdc25C expression levels were significantly greater in cervical cancer patient group than in those of control group. Thus Cdc25 phosphatase might play an important role in carcinogenesis of cervical carcinoma. Further studies based on the correlation between Cdc25 phosphatase and survival rate would be need to support Cdc25 phosphatase as a prognostic factor of cervical carcinoma.
Subject(s)
Female , Humans , Blotting, Western , Carcinogenesis , cdc25 Phosphatases , Cell Cycle , Cervix Uteri , DNA , Flow Cytometry , Genital Diseases, Female , Lymph Nodes , Neoplasm Metastasis , RNA, Messenger , Survival Rate , Uterine Cervical NeoplasmsABSTRACT
Objective: To explore the inhibitory effect of artesunate(Art)on human esophageal carcinoma cells and to study the related mechanism.Methods: Nude mice were inoculated with Eca109 cells subcutaneously on the left upper limbs to establish esophageal carcinoma model.The model mice were divided into 5 groups: first group received 100 mg/kg Art,second group 200 mg/kg Art,third group 300 mg/kg Art,forth group 3 mg/kg cisplatin(DDP),and the fifth group received normal saline.Mass and volume changes of transplant tumors in different groups were observed.Flow cytometry was used to detect the cell cycle,apoptosis,and the expression of CDC25A protein,Smad3 protein and TGF-?protein in the transplanted tumors in mouse model.RT-PCR was used to detect the expression of CDC25A,Smad3 and TGF-?mRNA in the transplanted tumors.Results: Nude mouse model bearing human esophageal carcinoma was success- fully created.Compared with the control group,the volume and mass of transplant tumors in Art groups were significantly smaller(P