Clinical significance of p27 Kip1 expression in advanced ovarian cancer

Background: Ovarian cancer is the most common gynecological malignancy. In patients with advanced ovarian cancer, some biological parameters have prognostic implementations. P27kip1 is an inhibitor of a cycline-dependent kinase, its loss, can contribute to tumor progression. Objective: This study aimed to examine the importance of P27KIP1 protein in predicting the prognosis and response to neoadjuvant chemotherapy in patients with advanced ovarian epithelial cancer and to compare the outcomes of immunohistochemistry with Quantitative Real-time PCR. Patients and methods: We have studied P27KIP1expression by both immunohistochemistry and Quantitative Realtime PCR from 88 patients with advanced ovarian carcinomas undergone radical debulking surgery and received Paclitaxel followed by Cisplatin every 3 weeks for a total of 6 cycles. We also studied their association with both chemotherapy response and patient survival. Results: Nuclear expression of p27KIP1 protein was intense in 86 normal ovarian tissues and 42 of 88 carcinomas. The P27kip1mRNA expression level by qRT-PCR was very low in ovarian cancer tissues relative to its adjacent normal tissues. The results were statistically significant by both methods of determination. p27KIP1 expression was significantly related to good prognostic parameters as low stage tumors, differentiated tumors, absence of ascites, residual disease < 2 cm, and response to chemotherapy but not with histopathological type in case of determination by immunohistochemistry. Comparison of P27kip1 by both immunohistochemistry and qRTPCR with different prognostic parameters revealed no significant difference between both methods in the assessment of these parameters. In 4 years of follow-up, 20.5% of patients were alive without evidence of disease. 6.8% were alive with disease. The disease-related four -year survival rate for the whole group was 28.2%. In multivariate analysis, residual disease, histological type, tumor differentiation, ascites was of independent prognostic significance. Conclusion: In ovarian cancer, patients with loss of p27KIP1 expression are at a greater likelihood of disease progression, p27KIP1 may be used as a molecular marker to predict response to chemotherapy and prognosis. Both immunohistochemistry and qRT-PCR have equal reliability in the determination of p27 KIP1

The expression and comparison of p57kip2 and p27kip1 in hydatidiform moles / 国际检验医学杂志

Objective To observe the level of imprinting gene p57kip2 and p27kip1 expression in different hydatidiform moles.To investigate the value of combined detection of p57kip2 and p27kip1 in diagnosis and differential diagnosis of hydatidiform moles.Methods We examined the immunohistochemical staining of p57kip2 and p27kip1 in 30 cases of complete hydatidiform moles and 86 cases of partial hydatidiform moles and 30 cases normal placenta,and also analyze the differences and correlation between the two genes of p57kip2 and p27kip1 in two different patterns of hydatidiform moles.Results The rate of expression of p57kip2 and p27kip1 in complete hydatidiform moles was obviously lower than that in partial hydatidiform moles and nor-mal placenta.There were significant differences in the expression of p57kip2 and p27kip1 among complete hy-datidiform moles,partial hydatidiform moles and normal placenta(P<0.05).p57kip2 had positive correlation with p27kip1 in complete hydatidiform moles(r=0.750,P<0.01),meanwhile there was negative correlation between p57kip2 and p27kip1 in partial hydatidiform moles(r= -1.000,P<0.01).Conclusion The differen-tial expression of p57kip2 and p27kip1 in complete hydatidiform moles and partial hydatidiform moles can be of certain value in the differential diagnosis of hydatidiform moles.Meanwhile,the combined methed is useful to the identification and classification of hydatidiform moles.

Biological characteristics of and protein expressions of p27kip1/PTEN in bone marrow mesenchymal stem cells from patients with systemic lupus erythematosus / 中华皮肤科杂志

Objective To evaluate biological behaviors of bone marrow mesenchymal stem cells (BMSCs) from patients with systemic lupus erythematosus (SLE),to confirm that the BMSCs are aging stems cells,and to explore mechanisms underlying their aging.Methods BMSCs were isolated from bone marrow of 6 patients with SLE (patient group) and 8 healthy human controls (control group) by density-gradient centrifugation and plastic adherence,and cultured in vitro.Optical microscopy was conducted to observe morphological changes and growth of BMSCs,and growth curves were drawn.The differentiation ability of BMSCs was evaluated through culture of them with adipogenic and osteogenic induction medium.Flow cytometry was performed to identify cellular surface markers and to analyze cell cycle and apoptosis,and scratch assay to assess the migration ability of BMSCs.Immunofluorcscence assay and Western blot analysis were carried out to analyze the distribution and expression of p27kip1/PTEN in BMSCs respectively.Results Patient-derived BMSCs,which had a broad,fiat and polygonal shape,showed decreased growth rate,migration activity as well as adipogenic and osteogenic ability compared with those from the controls.There was a significant increase in the proportion of BMSCs in early stage (17.98％ ± 3.26％ vs.8.23％ ± 3.25％,t =3.91,P =0.011) as well as in middle to late stages (16.80％ ± 9.63％ vs.3.33％ ± 2.21％,t =2.99,P=0.048) of apoptosis in the patient group compared with the control group.Moreover,compared with the control group,the patient group showed a significantly higher proportion of BMSCs arrested in the G0/G1 phase (92.34％ ± 5.80％ vs.78.65％ ± 3.22％,t =3.635,P =0.015),but a lower proportion in the S phase (0.86％ ± 1.72％ vs.5.06％ ± 1.874％,t =3.084,P=0.027).The protein expressions of p27 and PTEN were significantly higher in the patient group than in the control group (p27/β-actin:t =2.784,P=0.039;PTEN/β-actin:t=4.812,P =0.041).Conclusion The BMSCs from SLE patients exhibit senescence-related features,which may be associated with elevated expression levels of p27kip1/PTEN.

Astaxanthin Inhibits Proliferation of Human Gastric Cancer Cell Lines by Interrupting Cell Cycle Progression

BACKGROUND/AIMS: Astaxanthin is a carotenoid pigment that has antioxidant, antitumoral, and anti-inflammatory properties. In this in vitro study, we investigated the mechanism of anticancer effects of astaxanthin in gastric carcinoma cell lines. METHODS: The human gastric adenocarcinoma cell lines AGS, KATO-III, MKN-45, and SNU-1 were treated with various concentrations of astaxanthin. A cell viability test, cell cycle analysis, and immunoblotting were performed. RESULTS: The viability of each cancer cell line was suppressed by astaxanthin in a dose-dependent manner with significantly decreased proliferation in KATO-III and SNU-1 cells. Astaxanthin increased the number of cells in the G0/G1 phase but reduced the proportion of S phase KATO-III and SNU-1 cells. Phosphorylated extracellular signal-regulated kinase (ERK) was decreased in an inverse dose-dependent correlation with astaxanthin concentration, and the expression of p27(kip-1) increased the KATO-III and SNU-1 cell lines in an astaxanthin dose-dependent manner. CONCLUSIONS: Astaxanthin inhibits proliferation by interrupting cell cycle progression in KATO-III and SNU-1 gastric cancer cells. This may be caused by the inhibition of the phosphorylation of ERK and the enhanced expression of p27(kip-1).

The expression of OCT4 and P27kip1 in renal hyaline cell carcinoma and their correlations / 天津医药

Objective To investigate the expressions of OCT4 and P27kip1 and their correlations in renal hyaline cell carcinoma. Methods A total of 24 samples of renal hyaline cell carcinoma were detected by immunohistochemistry. The rate of cells with positive labelling of OCT4 and P27kip1 and their intra cellular distribution were observed in renal hyaline cell carcinoma. The expression levels of OCT4 and P27kip1 were compared between different gender, age (<60y and≥60y) and cancer cell metastasis groups. Results The rates of cells with positive OCT4 and P27kip1 expressions were 66.7%and 75% in renal hyaline cell carcinoma respectively. The ratio of cells with high, middle and low expression of OCT 4 were 33.3%, 20.5%and 12.5%in renal hyaline cell carcinoma tissues. OCT4 was mainly expressed in cytoplasm and nuclei with a few was expressed in the cell membrane. The ratio of cells with high, middle and low expression of P27kip1 were 12.5%, 25%and 37.5%in renal hyaline cell carcinoma tissues respectively. The positive staining of P27kip1 was in the cytoplasm and cell membrane, and a small number of nuclei were expressed. There were no significant differences in OCT4 and P27kip1 between different age and gender groups. There were significant differences in OCT4 and P27kip1 expressions between patients with metastasis and patients without metastasis (P < 0.05). A negative correlation was found between OCT4 and P27kip1 expres?sions in renal hyaline cell carcinoma (rs=-0.408, P<0.05). Conclusion The expression of OCT4 is negatively correlated with the expression of P27kip1. Inhibiting expressions of OCT4 or P27kip1, especially knocking down P27kip1 in cytoplasm can be used as a new diagnosis and treatment of renal cancer gene therapy.

Effects of epidermal growth factor on cell cycle and cell cycle-related regulatory factors of human esophageal squamous cell carcinoma cell line Eca109 / 中国免疫学杂志

Objective:To investigate the effects of epidermal growth factor (EGF)on cell cycle and cell cycle-related regulatory factors of human esophageal squamous cell carcinoma (ESCC) cell line Eca109.Methods: Serum starved Eca109 cells were treated with 20 ng/ml recombinant human EGF(rhEGF)for 24 h.The cell cycle phase distribution was detected by flow cytometry.The mRNA and protein expression levels of p21CIP1/WAF1(p21) and p27KIP1(p27) were detected by real-time quantitative reverse transcription polymerase chain reaction(qRT-PCR)and Western blot,respectively.Results: The proportions of G1 phase cells in EGF group and control group were ( 54.90 ±0.82 )% and ( 65.94 ±0.74 )%.The mRNA and protein expression levels of p 21 in EGF group was significantly higher ,and p27 was significantly lower than that in control group ( P<0.01 ) .Conclusion: EGF facilitates G1-S phase transition,and promotes the proliferation of Eca 109 cells,which may be associated with the up-regulation of p21 and down-regulation of p27.

Effect of Skp2 antisense oligodeoxynucleotide on growth and proliferation of colorectal cancer SW 480 cells / 重庆医学

Objective To investigate the effect of S-phase kinase-associated protein 2 antisense oligodeoxynucleotide (Skp2 ASODN) on the growth and proliferation of colorectal cancer cells SW480 and its mechanism .Methods The liposome-mediated dif-ferent concentrations of Skp2 ASODN was adopted to transfect SW480 cells ,the final concentrations of 0 .050 ,0 .125 ,0 .250 ,0 .500 , 1 .000 ,2 .000 ,4 .000 μmol/L were respectively set up with Skp2 nonsense oligodeoxynucleotides(NSODN) and blank group as con-trol .Then the inhibited effect of growth and proliferation of colorectal cancer cells were measured by the inverted microscope obser-vationandmethylthiazolyltetrazolium(MTT),thecellcyclewassurveyedbytheflowcytometry(FCM).TheexpressionofmRNA and protein of Skp2 and P27kip1 were inspected by reverse transcription-polymerase chain reaction(RT-PCR) and immunocytochem-istry methods .Results The inverted microscope observed that the SW480 cells had no change in form and grew at a slower speed . When the Skp2 ASODN concentration reached 0 .125 μmol/L ,the inhibition rate was 14 .48% (P0 .05) ,but its protein expression was elevates obviously (P<0 .01) .Conclusion Skp2 ASODN can inhibit the growth and proliferation of SW480 cells .Its possible mechanism is that the ubiq-uitin degradation of the Skp2 to P27kip1 is decreased by the gene occlusion effect of Skp2 ASODN ,thus the cell cycle progression is arrested .

Silencing of KIF14 interferes with cell cycle progression and cytokinesis by blocking the p27(Kip1) ubiquitination pathway in hepatocellular carcinoma

Although it has been suggested that kinesin family member 14 (KIF14) has oncogenic potential in various cancers, including hepatocellular carcinoma (HCC), the molecular mechanism of this potential remains unknown. We aimed to elucidate the role of KIF14 in hepatocarcinogenesis by knocking down KIF14 in HCC cells that overexpressed KIF14. After KIF14 knockdown, changes in tumor cell growth, cell cycle and cytokinesis were examined. We also examined cell cycle regulatory molecules and upstream Skp1/Cul1/F-box (SCF) complex molecules. Knockdown of KIF14 resulted in suppression of cell proliferation and failure of cytokinesis, whereas KIF14 overexpression increased cell proliferation. In KIF14-silenced cells, the levels of cyclins E1, D1 and B1 were profoundly decreased compared with control cells. Of the cyclin-dependent kinase inhibitors, the p27Kip1 protein level specifically increased after KIF14 knockdown. The increase in p27Kip1 was not due to elevation of its mRNA level, but was due to inhibition of the proteasome-dependent degradation pathway. To explore the pathway upstream of this event, we measured the levels of SCF complex molecules, including Skp1, Skp2, Cul1, Roc1 and Cks1. The levels of Skp2 and its cofactor Cks1 decreased in the KIF14 knockdown cells where p27Kip1 accumulated. Overexpression of Skp2 in the KIF14 knockdown cells attenuated the failure of cytokinesis. On the basis of these results, we postulate that KIF14 knockdown downregulates the expression of Skp2 and Cks1, which target p27Kip1 for degradation by the 26S proteasome, leading to accumulation of p27Kip1. The downregulation of Skp2 and Cks1 also resulted in cytokinesis failure, which may inhibit tumor growth. To the best of our knowledge, this is the first report that has identified the molecular target and oncogenic effect of KIF14 in HCC.

Clinical significance and expression of Skp2, p27 and p21 in Ovarian Serous Cystadenocarcinomas / 中国综合临床

ObjectiveTo investigate the correlation of Skp2,p27kiP1 and p21WAF1 expression with the clinicopathological features of ovarian serous cystadenocarcinomas.Methods Expressions of Skp2 ,p27kiP1 and p21WAF1 were examined by immunohistochemical staining in 124 epithelial ovarian tumors (25 serous cystadenomas, 19 borderline serous cystadenomas, and 80 serous cystadenocarcinomas) Results(1) The expression of Skp2 in serous cystadenocarcinomas (47.5％)was significantly higher than that in borderline serous cystadenomas (0％)and serous cystadenomas (0％)(P ＜ 0.001) .The p27kiP1 expression in serous cystadenocarcinomas (35.0％) was significantly lower than that in borderline serous cystadenomas(73.7％)and serous cystadenomas (80.0％) .The p21WAF1 staining frequency in serous cystadenocarcinomas (38.8％)was significantly lower than in borderline serous cystadenomas (73.7％)and serous cystadenomas (80.0％) .(2) The Skp2 protein expression in serous cystadenocarcinomas was positively correlated with clinicopathological stage,histological differentiation degree and lymph node metastasis of the tumors.The p27kiP1, p21WAF1 protein expression in serous cystadenocarcinomas was reversely correlated with clinicopathological stage and histological differentiation degree of the tumors(Ps ＜ 0.05) .(3) The Skp2 protein expression in serous cystadenocarcinomas was reversely correlated with that of p27kiP1 , p21WAF1.Conclusion The Skp2 protein expression in serous cystadenocarcinomas was increased and positively correlated with the clinicopathological features of ovarian serous cystadenocarcinomas.Skp2 protein expression was reversely correlated with p27kip1 ,p21WAF1.Skp2 protein expression may play an important role in the development and progression of serous cystadenocarcinomas.

Expression of Foxo3a and p27kip1 in lumbar dorsal root ganglia (DRG) after rat sciatic nerve injury / 中华显微外科杂志

Objective To investigate the expression of Foxo3a and p27kip1 in lumbar dorsal root ganglia (DRG) after injury of sciatic nerve in rats. Methods Adult rats were randomly divided into control group and experimental group. The rats in experimental group were subjected to sciatic nerve clamp.Expression and distribution of Foxo3a and p27kip1 and cellular proliferation and axon regeneration in DRG was detected by western blot and immunohistochemistry. Results Foxo3a protein levels begined to reduce at 1 day (7.0 ± 3.5), reached valley at 2 day (6.0 ± 3.8) after injury, and following Foxo3a downregulation, p27kip1 protein levels begined to decrease at 2 day (29.0 ± 3.5), reached valley at 7 day (21.0± 3.0) after injury. Down-regulation of Foxo3a and p27kip1 was expressed predominantly in neurons and glial cells by double immunolabelling. Foxo3a and p27kip1 were expressed in neurons [(37.8 ± 5.7)%, (43.3 ±4.3)%] and glial [(22.4 ± 3.9)%, (13.8 ± 3.2)%] cells in sections of DRG at 2 day after injury less than neurons [(73.6 ± 2.5)%, (84.1 ± 3.7)%] and glial [(61.3 ± 4.4)%, (68.7 ± 5.6)%] cells in sections of normal DRG. Proliferating cell nuclear antigen (PCNA) and GAP-43 were up-regulation from 2 day, and PCNA reached peak at 7 day after injury.The glial cells were the main type of cellular proliferation.Conclusion Down-regulation of Foxo3a and p27kip1 in lumbar DRG is correlated with the proliferation of glial cells and axonal regeneration after sciatic nerve injury.

Effects of triptolide on cell cycle and expression of P21wap1/cip1 and P27kip1 in human multiple myeloma RPMI 8226 cells / 中草药

Objective: To investigate the effect of triptolide on cell proliferation, cell cycle distribution, DNA and protein expression regulation of P21wap1/cip1 and P27kip1 in human multiple myeloma RPMI 8226 cells. Methods: Cell viability was detected by MTT assay and cell cycle distribution was measured by flow cytometry. Effect on mRNA expression of P21wap1/cip1 and P27kip1 was detected by RT-PCR. The change of protein expression was measured by Western blotting. Results: Triptolide of varying concentration significantly induced the inhibition of proliferation in a dose-dependent manner and G0/G 1 phase arrest of the cell cycle progression. The events were coincided with the upregulation of the mRNA and protein expression of P21wap1/cip1 and P27kip1. Conclusion: These results suggest that triptolide might exhibit its strong antitumor effect via alternation of P21wap1/cip1 and P27kip1. It provides framework for a clinical evaluation of triptolide.

Mechanism of cyclin-dependent inhibitor p27~(Kip1) in regulating the differentiation of immortalized human neural progenitor cells / 解剖学报

Objective To investigate whether there is any functional link between p27~(Kip1) function and all-trans retinoic acid (RA) in the control of neuronal differentiation of immortalized human neural progenitor cells (hSN12W-TERT cells). To investigate the mechanism by which p27~(Kip1) regulates the differentiation of immortalized human neural progenitor cells. Methods hSN12W-TERT cells were derived from the striatums of human embryos at 12 weeks gestation and cultured with serum-free medium in presence of EGF and bFGF. At the appropriated time, hSN12W-TERT cells were exposed to 1μmol/L RA for 3, 5, 7 days respectively. The experiment was repeated there times. Cell cycle analysis was performed by flow cytometry analysis (FACS). The expression of p27~(Kip1), p21~(cip1), cyclin-dependent kinase 2 (cdk2), p-cdk2 and S-phase kinase-associated protein 2 (skp2) in hSN12W-TERT cells before and after RA treatment cells were determined by using Western blotting analysis. Results FACS result showed that 77.25% of proliferating hSN12W-TERT cells were in the G1/G0-phase while 9.38% of cells in the S-phase. Following RA treatment, cell growth was arrested, and 85.68% of cells accumulated in G1/G0-phase while 8.57% of cells in the S-phase. Western blotting analysis demonstrated that the levels of p27~(Kip1) in the hSN12W-TERT cells increased following 3 days' treatment with RA compared with those of normal untreated cells, with a peak at 5 days (P<0.05). The similar results were acquired both in nuclear proteins and in cytoplasm proteins of hSN12W-TERT cells. The expression level of p21~(cip1) decreased in response to RA treatment. RA did not affect the expression of cdk2, but the expression of p-cdk2, which represented the activity of cdk2, was markedly decreased in response to RA treatment. Skp2, which was required for the ubiquitin-mediated degradation of p27~(Kip1), was detected in proliferating hSN12W-TERT cells. The expression of skp2 reduced dramatically in response to RA treatment in a time-dependent manner.Conclusion There is a functional link between RA and p27~(Kip1) function in the control of neuronal differentiation in hSN12W-TERT cells. P27~(Kip1) plays a key role during neuronal differentiation. Moreover, high levels of p27~(Kip1) are associated with its degradation inhibiting through reducing proteasome-dependent proteolysis.

Proapoptotic mechanism and changes of endogenous TGF-β_1 in NB4 cells induced by exogenous TGF-β_1 / 中国病理生理杂志

AIM:To study the effects of transforming growth factor-β_1 (TGF-β_1) on cell apoptosis,cell cycle,production of endogenous TGF-β_1,expressions of P27~(Kip1),cyclin E and bcl-2 mRNA levels in NB4 cells. METHODS:Apoptotic morphological changes were observed by Wright-Giemsa staining. Cell cycle and apoptosis were detected with flow cytometry. Semiquantitative RT-PCR was used to examine the mRNA levels of endogenous TGF-β_1,P27~(Kip1),cyclin E and bcl-2. RESULTS:TGF-β_1 significantly restrained the growth and promoted the apoptosis of NB4 cells. The blockage of NB4 cells treated by TGF-β_1 at concentration of 5 μg/L was in G1 phase. Endogenous TGF-β_1 mRNA expression in NB4 cells was up-regulated when the concentration of exogenous TGF-β_1 was <5 μg/L. Meanwhile,the expression of endogenous TGF-β_1 mRNA was down-regulated when the concentration of exogenous TGF-β_1 was 10 μg/L. After treated with TGF-β_1 at concentration of 5 μg/L,P27~(Kip1) mRNA expression in NB4 cells was up-regulated,cyclin E and bcl-2 were reduced. CONCLUSION:TGF-β_1 is able to induce apoptosis and cell cycle distribution abnormally in NB4 cells by (1) Up-regulation of endogenous TGF-β_1,so that NB4 cells was induced into apoptosis through consequently high expression of P27~(Kip1). (2) TGF-β_1 may lead to cell cycle arrest by inhibiting the expression of cyclin E directly,or by inhibiting the activity of cyclin E through the increased expression of P27~(Kip1). (3) Down-regulation of bcl-2 induces apoptosis of NB4 cells.

Significance of p27(kip1) as potential biomarker for intracellular oxidative status

Our previous proteomic study demonstrated that oxidative stress and antioxidant delphinidin regulated the cellular level of p27(kip1) (referred to as p27) as well as some heat shock proteins in human colon cancer HT 29 cells. Current study was conducted to validate and confirm the regulation of these proteins using both in vitro and in vivo systems. The level of p27 was decreased by hydrogen peroxide in a dose-dependent manner in human colon carcinoma HCT 116 (p53-positive) cells while it was increased upon exposure to hydrogen peroxide in HT 29 (p53-negative) cells. However, high concentration of hydrogen peroxide (100 micrometer) downregulated p27 in both cell lines, but delphindin, one of antioxidative anthocyanins, enhanced the level of p27 suppressed by 100 micrometer hydrogen peroxide. ICR mice were injected with varying concentrations of hydrogen peroxide, delphinidin and both. Western blot analysis for the mouse large intestinal tissue showed that the expression of p27 was upregulated by 25 mg/kg BW hydrogen peroxide. To investigate the association of p27 regulation with hypoxia-inducible factor 1-beta (HIF-1beta), the level of p27 was analyzed in wild-type mouse hepatoma hepa1c1c7 and Aryl Hydrocarbon Nuclear Translocator (arnt, HIF-1beta)-defective mutant BPRc1 cells in the absence and presence of hydrogen peroxide and delphinidin. While the level of p27 was responsive to hydrogen peroxide and delphinidin, it remained unchanged in BPRc1, suggesting that the regulation of p27 requires functional HIF-1beta. We also found that hydrogen peroxide and delphinidin affected PI3K/Akt/mTOR signaling pathway which is one of upstream regulators of HIFs. In conclusion, hydrogen peroxide and antioxidant delphinidin seem to regulate intracellular level of p27 through regulating HIF-1 level which is, in turn, governed by its upstream regulators comprising of PI3K/Akt/mTOR signaling pathway. The results should also encourage further study for the potential of p27 as a biomarker for intracellular oxidative or antioxidant status.

Impact of 12-lipoxygenase on p27Kip1 expression in experimental diabetic glomeruli / 中华肾脏病杂志

Objective To investigate the effect of 12-lipoxygenase(12-LO) on the p27Kip1 expression in diabetic glomeruli. Methods Mesangial cells were exposed to 12-LO product 12 (S)-HETE (10-7 mmol/L) with or without p38 MAPK (p38) inhibitor (SB203580, 1 μmol/L) for 24 hours. Rats fed with high fat diet received low dose streptozotoein (ST-Z, 35 mg/kg, IP injection) to develop type 2 diabetes and were divided into 2 groups: low dose STZ, low dose STZ+12-LO inhibitor cinnamyl-3,4-dihydroxy-α-cynanocinnamate (CDC, 8 mg/kg) treatment. Rats fed with regular chow were divided into two groups: controls, CDC treatment. The rats received injection of CDC or vehicle subcutaneously in the hind leg. CDC or vehicle injection was performed three times weekly on alternate days. All the rats were sacrificed after 4 weeks, Wild type and 12-LO knockout C57BL/6 mice were divided into 4 groups: wild type control, 12-LO knockout, STZ-induced wild type type 1 diabetes and STZ-induced 12-LO knockout type 1 diabetes. All the mice were sacrificed after 16 weeks. Urine, blood, kidney cortical tissue and isolated glomeruli by sieving method were collected at the end of study respectively. Western blot and immunohistochemistry for target protein were performed respectively. Results Inhibition of p38 activation could significantly reduce p27Kip1 expression induced by 12 (S)-HETE in mesangial cells (P<0.01). Increased glomerular volume, microalbuminuria, elevated glomeluli p38 activation, p27Kip1 expresssion in type 2 diabetic glomeruli was decreased after CDC treatment (P<0.01). Compared with wild type diabetic mice, glomerular p38 activation, p27Kip1 exprcsssion and extracellular matrix accumulation in the 12-LO knockout diabetic mice were significantly decreased (P <0.01, respectively). Conclusions 12-LO induces p27kipl expression via p38 pathway in diabetic glomeruli.

Expression and significance of p27kip1, CyclinD1, Cyclin E in transitional cell carcinoma of bladder / 肿瘤研究与临床

Objective To investigate the expression of p27kip1, CyclinE, CyclinD1 in bladder transitional cell carcinoma (BTCC), and evaluate their relationship with turnout cell proliferation. Methods To study the expression of p27kip1, CyclinE and CyclinD1 protein in 41 cases of paraffin-embedded BTCC tissue and in 12 cases of cystitis tissues by immunohistochemical method SP. Results There was a negative relationship between p27kip1 expression and clinical stage and pathological grade.There was no relationship with lymph node metastasis (P>0.05). There was a positive relationship between CyclinE expression and clinical stage, pathological grade and lymph node metastasis (P<0.05). There was no relationship between CyclinD1 expression and clinical stage, pathological grade and lymph node metastasis (P>0.05). Conclusion The results of this study showed that the abnormal expression of p27kip1, CyclinE, may play important roles in the genesis and progression of BTCC. CyclinD1 may take activation in the early term of BTCC.

p27~(kip1) Expression in Radio-resistance of Esophageal Cancer Cells / 华中科技大学学报(医学版)

Objective To investigate the relationship between the p27~(kip1) expression and the change of radiosensitivity of esophageal cancer cells.Methods Radio-resistant cells (EC9706R) were gradually isolated by means of repeated gamma-ray irradiation (6 000 cGy in total) upon human squamous esophageal carcinoma cells (EC9706).The radiosensitivity of these two types of cells was measured by standard colony formation assays,their cell cycle distribution analyzed by flow cytometry respectively,and the p27~(kip1) expression detected by immunocytochemistry.Results As compared with their parent cells,the isolated human squamous esophageal carcinoma cells(EC9706) showed clearly greater radio-resistance(for EC9706R,SF_2=65.71%,D_0=2.20 Gy,D_q=1.61 Gy,N=2.07,and for EC9706,SF_2=46.72％,D_0=1.61 Gy,D_q=0.99 Gy,N=1.85;SF_2,D_0,Dq and N were all higher).As to the cell cycle distribution,the population of G_1 phase cells in EC9706R cells was significantly decreased,but the proportion of S phase cells was significantly increased as compared with the parent cells.Immunocytochemistry revealed that the p27~(kip1) expression of EC9706R cells was clearly lower than that of EC9706 cells.Conclusion Cell phase change due to the decrease of p27~(kip1) expression may be one of the mechanisms of the generation of radio-resistance in esophageal cancer cells.

Effect of 12(S)-HETE on the p27~(kip1) expression in rat mesangial cells and glomeruli / 中国免疫学杂志

Objective:To investigate the effect of 12(S)-HETE on the p27~(kip1) expression in mesangial cells and glomeruli.Methods:Mesangial cells were exposed to 12(S)-HETE.12(S)-HETE was infused to rats by osmotic mini-pump.Total protein content measurement for cell hypertrophy,RT-PCR for mRNA expression and Western blot for protein expression were performed respectively.Results:12(S)-HETE stimulation induced mesangial cell hypertrophy and p27~(kip1) protein expression,but not p27~(kip1) mRNA expression.Furthermore,p27~(kip1) mRNA and protein expression in the glomeruli were significantly increased by 12(S)-HETE stimulation using osmotic mini-pump.Conclusion:12(S)-HETE plays an important role in the pathogenesis of glomerular cell hypertrophy and senescence through upregulation of p27~(kip1) expression.

Expression Profiles of Retinoblastoma Protein and p27Kip1 According to HER2 Amplification Status in Primary Breast Cancer / 한국유방암학회지

PURPOSE: Human epidermal growth factor receptor-2 (HER-2)/neu amplification affects the cell proliferation through the modulation of multiple G1 cell cycle regulators in breast tumor cells. We performed this study to investigate whether retinoblastoma protein (pRB) and p27Kip1 were differently expressed according to the HER2 amplification status in human breast cancer. METHODS: HER2 amplification was assayed by fluorescence in situ hybridization and the expression of cell cycle regulators were assayed by immunohistochemistry on 153 consecutive invasive breast cancers. The proliferative activity of breast cancer was analyzed according to the HER2 amplification and cell cycle protein expression status. RESULTS: HER2 amplification was observed in 39 (25.5%) of 153 breast cancers. In the HER2 amplified breast cancers, the pRB expression was significantly increased (p=0.011) whereas there was no significant relationship between HER2 amplification and p27Kip1 expression. There was an inverse correlation between pRB expression and Ki-67 labeling index in the HER2 amplified breast cancers (p=0.036). In contrast, Ki67 labeling index was significantly decreased as p27Kip1 expression increased in HER2 non-amplified breast cancers (p=0.028). In HER2 non-amplified breast cancers, we could not observe any association between the pRB expression and Ki67 labeling index. CONCLUSION: The proliferation of the breast cancers was associated with pRB expression in HER2 amplified tumors whereas it was associated with p27Kip1 expression in HER2 non-amplified tumors. The results of the current study indicate that the cell proliferative activity of the breast cancer is under different growth signal pathways according to HER2 amplification status.

Significance of Notch Expression in Acute Myeloid Leukemia / 대한혈액학회지

BACKGROUND: Notch is a gene family encoding receptors to transduce intercellular signals involved in cell-fate determination. Although several lines of evidence indicate that abnormal Notch signaling may contribute to neoplastic transformation, little is known regarding the role of Notch in the pathogenesis of leukemia. METHODS: To explore the functional significance of Notch1 in acute myeloid leukemia (AML), the expression of Notch1 and its association with survivin and p27Kip1 expression was examined in 50 patients with de novo AML. RESULTS: Notch1 transcripts were expressed in 40 (80%) cases with a variable degree of expression, and the fraction of AML cells in the G0/G1 phase was higher in Notch1-positive cases than in Notch1-negative cases. Survivin was shown to be present in 38 (76.0%) cases, and Notch1 expression highly correlated with survivin mRNA expression (r=0.7170, P<0.001). p27Kip1 was present in 40 (80.0%) cases of AML and p27Kip1 expression was significantly associated with Notch1 expression (r=0.8770, P<0.001). Except for one case, the simultaneous expression of survivin and p27Kip1 was not seen in all cases that were negative for Notch1 expression. There were no differences in clinical outcomes according to Notch1 expression. CONCLUSION: Notch1 expression was a frequent event and has functional significance in the alteration of the cell cycle in AML cells. Notch1 expression was also significantly associated with survivin and p27kip1 expression in AML cells. To evaluate the clinical significanceand functional role of Notch1 expression in the aberrant regulation of survivin and p27Kip1 expression in de novo AML, a further study with a larger number of patients is necessary.