ABSTRACT
@#[Abstract] Objective: To investigate the expression of wild type p53 induced phosphatase 1 (Wip1) in small cell lung cancer (SCLC) cells and the serum of SCLC patient and its relationship with clinical prognosis. Methods: Real time quantitative PCR (qPCR) was used to detect the expression of Wip1 in SCLC cells and serum samples. Results: The expression of Wip1 in drug-resistant SCLC cells was significantly higher than that in sensitive cell lines (P<0.01). The expression of Wip1 in serum of SCLC group was significantly higher than that of normal control group (P<0.05); the expression of Wip1 in serum of patients with chemotherapy resistance was significantly higher than that in patients with chemotherapy sensitivity (all P<0.05); the serum Wip1 level was correlated with disease stage, chemotherapy sensitivity and survival status of SCLC patients (all P<0.05). The area under ROC curve of Wip1 predicting the prognosis of SCLC was 0.836 (95%CI:0.8230-0.9600, P<0.01); the expression lever of Wip1 was significantly correlated with progression free survival and overall survival time of SCLC patients (all P<0.05). Disease stage, chemosensitivity and Wip1 expression were independent prognostic factors for SCLC patients (all P<0.05). Conclusion: The expression of Wip1 in serum of SCLC patients may be related to chemotherapy sensitivity and prognosis. Wip1 may be a potential biomarker for therapeutic efficacy and prognosis evaluation of SCLC patients.
ABSTRACT
Wildtype p53-induced phosphatase 1 (Wip1) is a serine/threonine protein phosphatase of 605 ami-no acids, which is expressed at high levels in many organs and tissues .As Wip1 is overexpressed in human tumors , analy-sis of Wip1 has focused primarily on its role in tumorigenesis .In recent years , it has also been shown that Wip 1 plays an important role in several physiological processes including adult neurogenesis , senescence , immunodeficiency and metabolic diseases.This review addresses how Wip1 participates in physiological and pathological conditions at cellular and molecular levels.
ABSTRACT
Objective To construct the Wip1 gene recombinant lentiviral expression vector and to investigate its effects on breast cancer cell biological behaviors .Methods Wip1 gene short hairpin RNAs (shRNA) was transfected into breast cancer MCF-7 cells through lentiviral infection method .Wip1 mRNA and protein expressions before and after transfection were detected by u-sing qRT-PCR and Western blotting .The effects of Wip1-shRNA on the proliferation ,apoptosis ,cell cycle ,invasion and metastasis in MCF-7 cells were determined by using the MTT assay ,flow cytometry and transwell invasion test .Interfering RNA molecule p53dsRNA inhibiting p53 gene expression (p53 dsRNA) was screened ,and the effect of p53 inhibition on MCF-7 cells invasion and metastasis was analyzed .Results After transfection for 48 h ,the cellular fluorescence in the Wip1-shRNA group was stronger , while which in the NC-shRNA group was weaker .Cellular Wip1 mRNA and protein expressions in the Wip1 shRNA group were 0 .291 ± 0 .025 and 0 .203 ± 0 .021 respectively ,which in the NC-shRNA group were 0 .954 ± 0 .090 and 0 .963 ± 0 .092 respectively , the difference between the two groups was statistically significant (P<0 .05) .The cellular survival rate at various time points had statistical difference between the two groups (P<0 .05) .The early and late cell apoptosis number in the NC-shRNA group was less than that in the Wip1-shRNA group ,the difference was statistically significant (P<0 .05) .The cells numbers at phase G0 +G1 and phase S in the NC-shRNA group were 53 .5 ± 3 .6 and 27 .3 ± 1 .5 respectively ,which in the Wip-shRNA group were 72 .3 ± 5 .2 and 14 .6 ± 0 .8 respectively ,the difference was statistically significant(P<0 .05) .The Transwell invasion and metastasis results showed that the cell transmembrane number in the NC-shRNA group was more than that in the Wip1-shRNA group(P<0 .05) .The cellu-lar p53 mRNA and protein expression had statistical difference between the control group and p53dsRNA group(P<0 .05) .Conclu-sion RNA interference can effectively suppress Wip1 expression in MCF-7 cells .Wip1 may affect the proliferation ,apoptosis ,cell cycle ,invasion and metastasis of breast cancer cells by modulating protein expression .p53dsRNA increases the invasion and metas-tasis ability of breast cancer MCF-7 cells by interfering p53 gene down-regulation .
ABSTRACT
Objective To investigate the related factors of clinical stage and prognosis in the patients with endometrial carcinoma and their relation with proto-oncogene Wip1 expression level.Methods The paraffin samples of resected endometrial carcinoma in 120 cases of endometrial carcinoma in our hospital from January 2002 to January 2012 were collected as the experimental group,the samples were verified by pathology.Contemporaneous 120 samples of biopsy normal endometrial tissue served as the control group.The expression leve of Wipl were detected in turo groups.Results (1) In the Wip1 immunohistochemical staining results:Wip1 immunohistochemical staining was negative or weak in normal endometrial tissue cells,while showed pale yellow to yellowish-brown in endometrial cancer tissue.The positive expression rate of Wip1 protein in endometrial carcinoma tissue was 77.5%(93/120),which was higher than 22.5%(27/120) in normal endometrial tissue,the difference was statistically significant (P<0.05).(2)In the Western blot results of Wip1 protein in endometrial cancer tissue and normal endometrial tissue:the relative amount of Wip1 protein in endometrial carcinoma tissue was 0.635±0.023,which was significantly higher than 0.325±0.018 in normal endometrial tissue,the difference between the two groups was statistically significant (P<0.05).(3)In the real time quantitative qRT-PCR results of various samples:Wip1 mRNA expression level was higher than that in normal endometrial tissue,which were 0.628±0.053 and 0.191±0.009 respectively,the difference between the two groups was statistically significant(P<0.05).(4) The expression level of Wip1 had no correlation with age,estrogen and progesterone status,HER2,lymph node status and TNM stage,but had correlation with P53 expression level.Conclusion (1) The Wip1 expression amount is high in endometrial carcinoma and low in normal endometrial tissue.(2)The Wip1 expression level has no relation with age,estrogen and progesterone status,HER2,lymph node status and TNM stage,while has association with P53 expression level.
ABSTRACT
AIM:To observe the inhibitory effect of siRNA targeting to Wip1 gene on the Wip1 gene expression in the colon cancer cells and to investigate the influence of Wip1 gene silencing on the chemotherapy sensitivity of colon cancer cells.METHODS:Wip1-811 siRNA targeting to Wip1 gene was transfected into RKO colon cancer cells with high expression of Wip1 gene.The mRNA expression of Wip1 was measured by real-time PCR.The protein level of Wip1 was detected by Western blotting.The viability of RKO colon cancer cells was measured by MTS assay.The cell apoptosis and cell cycle were analyzed by flow cytometry.RESULTS: Wip1-811 siRNA efficiently inhibited the expression of Wip1 at mRNA and protein levels.The enhanced chemotherapy sensitivity of RKO colon cancer cells was observed after inhibition of Wip1 gene expression.The viability of RKO colon cancer cells was decreased from (89.4 ±6.6)%to (74.7 ±3.9)%af-ter treated with 5-fluorouracil (P<0.05) and decreased from (77.9 ±2.4)%to (66.7 ±2.9)%after treated with oxali-platin ( P<0.05 ) .The cell apoptotic rate was increased from ( 7.7 ±0.5 )% to ( 12.3 ±3.2 )% and from ( 14.7 ± 2.1)% to (34.0 ±2.1)% when RKO colon cancer cells were treated with 5-fluorouracil and oxaliplatin, respectively (P<0.05).CONCLUSION:Wip1 gene silencing enhances chemotherapy sensitivity of colon cancer cells.
ABSTRACT
Objective:To investigate the expression and clinical significance of wild-type p53-induced phosphatase 1 (Wip1) in thyroid carcinoma and biological effect of siRNA-targeting Wip1 on the thyroid carcinoma cell line. Methods:Immunohistochemistry and reverse transcription polymerase chain reaction (RT-PCR) were performed to detect the expression of Wip1 in 73 specimens of thy-roid carcinoma tissues and normal thyroid tissues (5 cm away from the margin of thyroid carcinoma), respectively. Wip1 siRNA was transiently transfected into the papillary thyroid carcinoma cell by using a liposome-mediated method and then detected by RT-PCR and Western blot. Methyl thiazolyl tetrazolium (MTT) assay and flow cytometry (FCM) were also conducted to observe cell proliferation, cell apoptosis, and cell cycle. Results:The positive rates of Wip1 protein were 80.8%in thyroid carcinoma tissues and 9.6%in the nor-mal tissues (χ2=47.036, P0.05). However, significant correla-tions among Wip1 expression, lymph node metastasis, clinical stages and tumor differentiation (P<0.05) were observed. RT-PCR and Western blot results showed that K1 cell-transfected Wip1 siRNA exhibited a relatively lower expression than normal cells (t=17.039, t=14.637, P<0.05). MTT assay results showed that the K1 cells transfected with Wip1 siRNA showed a lower survival fraction, higher cell apoptosis, higher percentage of G0/G1 phases, and lower cell concentration in G2/M and S phases (P<0.05). Conclusion:Wip1 pro-tein and mRNA were increased in thyroid carcinoma and are correlated with lymph node metastasis, clinical stages and tumor differenti-ation. Wip1 may be involved in proliferation, apoptosis, and cycle of thyroid cancer cells.
ABSTRACT
[Objective] The aim of this study was to establish a quantitative SYBR Green Ⅰ real-time PCR method for detection of wide-type p53-induced phosphatase 1 (Wip1 or PPM1D) gene expression level in non-small cell lung cancer (NSCLC),and to investigate the relationship between Wip1 mRNA expression level and the clinicopathological characters.[Method] Real-time PCR was employed to determine the expression level of Wip1 mRNA in 44 specimens of NSCLC tissues and their adjacent normal tissues.[Results] In the 44 specimens,the expression of Wip1 mRNA in both cancer tissues and adjacent normal lung tissues were positive.Wip1 gene was overexpressed in 17 specimens among 44 NSCLC specimens.The rate was 38.6%.The relative level of Wip1 mRNA in NSCLC tissues was significantly higher than the adjacent normal lung tissues (Ratio ＝ 2.1644 ± 1.394,P ＜ 0.01).The expression of Wip1 mRNA was also correlated with pathological staging (F ＝ 5.08,P ＝ 0.013).[Conclusion] The established SYBR Green Ⅰ quantitative real-time PCR method can successfully detect the expression level of Wip1 mRNA.The results suggested that Wip1 may be involved in the development of NSCLC.
ABSTRACT
Wip1 is a nuclear protein and a member of serine/threonine specific protein phosphatase type 2C(PP2C)family.It was initially identified as a gene whose expression was induced in response to ? or UV radiation in a p53-dependent manner and a negative feedback regulation of p38MAPK-p53 signaling.Then,Wip1 gene was confirmed a proto-oncogene and amplified or overexpressed in several human tumor types.This review will introduce the structures and functions of Wip1 and details on the signaling process of cancer progression.