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Objective:To examine the expression of programmed cell death 1 (PD-1) and its ligand (PD-L1) in epithelial ovarian cancer (EOC) tissues, and investigate the correlation among their expression, clinicopathological features and prognosis.Methods:The specimens of 180 patients with EOC treated in the First Affiliated Hospital of Dalian Medical University from October 2002 to December 2013 were confirmed by pathological examination. The pathological tissue specimens of subtypes ，included 120 cases of serous carcinoma, 30 cases of mucinous carcinoma, 20 cases of endometrioid carcinoma, and 20 cases of clear cell carcinoma. The normal paracancerous tissues of 50 cases randomly selected from the 180 patients as control group. Immunohistochemical SP method was used to detect the expressions of both PD-1 and PD-L1 in epithelial ovarian cancer tissues, and the relationships among their expressions,the clinicopathological parameters and prognosis were respectively analyzed.Results:（1） PD-1 was expressed in lymphocytes infiltrated in EOC tissues, and PD-L1 was expressed in the cell membranes of cancer tissues. In all EOC cases, 33 cases (18.3%, 33/180) of both PD-1 and PD-L1 were highly expressed, and only 1 (2.0%, 1/50) of control group showed high expression. There was statistically significant difference between two groups ( P<0.01). (2) Among the four subtypes tissue specimens of EOC, the high expression rate of PD-1 was 25.0% (30/120) for serous carcinoma, 3/15 for endometrioid carcinoma, 0 (0/30) for mucinous carcinoma, and 0 (0/15) for clear cell carcinoma. The high expression rate of PD-L1 was 23.3% (28/120) for serous carcinoma, 3.3% (1/30) for mucinous carcinoma, 2/15 for endometrioid carcinoma, and 2/15 for clear cell carcinoma. Both PD-1 and PD-L1 expressions in the four sub-types of tissue specimens were significantly different ( P<0.05). The high expression rate of both PD-1 and PD-L1 was 9.2% (8/87) in the early stage and 26.9% (25/93) in the late stage. There was a statistically significant difference between the two groups ( P<0.01). Similarly, the expression of both PD-1 and PD-L1 were significantly higher in the cases of high-grade EOC （type Ⅱ） than those of low-grade （type Ⅰ） and in the cases of EOC distributed bilaterally than that distributed unilaterally, and there were statistically significant differences ( P<0.05). (3) The Kaplan-Meier survival analysis showed that the survival time were respectively 35 and 36 months in the cases with high expressions of both PD-1 and PD-L1, and the survival time were the same as 61 months in the cases with low expression of both PD-1 and PD-L1, and the comparison was statistically significant ( P<0.05). Conclusions:The expression levels of PD-1 and PD-L1 in EOC tissues are higher than those in adjacent tissues, especially in serous carcinomas. The expression of both PD-1 and PD-L1 is higher in specimens of the patients with advanced stages. The results showed that the high expression of both PD-1 and PD-L1 is an indicator of poor prognosis of patients suffering from EOC.
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Objective To detect phosphorylated-extracellular signal-regulated kinase (p-ERK1/2) protein expression in epithelial ovarian cancer and cell lines,and to examine the effects of mitogen-activated protein kinase (MAPK) kinase (MEK) inhibitor AZD6244 on cell proliferation,apoptosis as well as cell cycle of ovarian cancer cells.To explore the function and significance of MAPK/extracellular signal-regulated kinase (ERK) signaling pathway in the development of ovarian cancer.Methods (1) A total of 104 cases of patients with ovarian cancer who accepted the treatment of gynecological surgery and being confirmed by pathological examination in First Affiliated Hospital,Dalian Medical University from January 2004 to December 2013 were selected.The expressions of p-ERK1/2 protein were detected by immunohistochemistry in ovarian cancer specimens,and the relationship between the expressions of p-ERK1/2 and the clinical features of patients was analyzed.(2) p-ERK1/2 and other related proteins were determined by western blot in various ovarian cancer cells,including SKOV3,OV2008,C13,A2780S,A2780CP,OVCAR4,OVCAR5,OVCAR8 and CAOV3 treated with or without MEK inhibitor.The cellular proliferation,apoptosis and cell cycle of ovarian cancer cells after treatment with MEK inhibitor were analyzed by methyl thiazolyl tetrazolium (MTT) assay and flow cytometry,respectively.Results (1) The immunohistochemical method showed that p-ERK1/2 between low grade serous carcinoma and clear cell carcinoma were not significantly higher expressed (P＞0.05).However,a lower level of the p-ERK1/2 expression were observed among high grade serous carcinoma,mucinous carcinoma and endometrioid carcinoma (all P＜0.05).There was no significant correlation between the protein expression of p-ERK1/2 and patients' age,pathological stage of surgery,and preoperative serum CA125 level (P＞0.05).(2) Western blot showed that the protein p-ERK1/2 was widely expressed in various ovarian cancer cell lines such as SKOV3,OV2008,C13,A2780S,A2780CP,OVCAR4,OVCAR5,OVCAR8 and CAOV3.After treatment with AZD6244 (5,10 μmol/L),the level of p-ERK 1/2 in OVCAR5 and OVCAR8 decreased significantly in dose-dependent manner.Additionally,we found a reduction of the expression level of cyclin D 1,caspase-3 and appeared cleaved poly adenosine diphosphate ribose polymerase (PARP) in OVCAR5 and OVCAR8,compared with control groups.MTT assays showed that OVCAR5,OVCAR8 and A2780S were differently inhibited in the dose-dependent manner after being treated with different concentrations of AZD6244 (0,2.5,5,10,25,50 and 100 μmol/L,all P＜0.05).Further tested by flow cytometry,the results showed that AZD6244 (5,10 μmol/L) was able to induce the apoptosis of OVCAR5,OVCAR8 and A2780S,as well as G0/G1 phase arrest,both in a dose-dependent manner (P＜0.05).Conclusions As the main active and functional unit of MAPK/ERK signaling pathway,p-ERK1/2 protein is expressed in both the tissues and various ovarian cancer cell lines.AZD6244 could down-regulated the expression of p-ERK1/2 in ovarian cancer cells,accompanied by the decreased proliferation and increased cell apoptosis of ovarian cancer cells.In conclusion,MAPK/ERK signaling pathway might play a role in the development and progression of ovarian cancer,and may be provide a novel option for molecular targeted therapies of the disease.
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Objective To examine the expressions of IKKε protein in the specimens and cells of epithelial ovarian cancer and investigate the effect of IKKε inhibitor on cell proliferation and apoptosis. Methods (1) A total of 118 cases of patients with the median age of 59 who have accepted surgical treatment due to ovarian cancer in the First Affiliated Hospital of Dalian Medical University from January 2006 to April 2013 were selected. Twenty cases of patients with the median age of 55 who have accepted hysterectomy and salpingo-oophorectomy due to uterine leiomyoma during the same period were selected as the control. The expressions of IKKε protein were detected by immunohistochemistry in normal ovarian tissues and epithelial ovarian cancer specimens,and the relationship between the expressions of IKKε and the clinical features of patients was analyzed. IKKε protein was determined by western blot in various ovarian cancer cells, including SKOV3, OV2008, C13, A2780S, A2780CP, OV4, OV5, OV8, and CAOV3 treated with or without IKKε inhibitor. The cellular proliferation and apoptosis of ovarian cancer cells after 48 hours treatment of IKKε inhibitor were analyzed by methyl thiazolyl tetrazolium (MTT) assay and flow cytometry, respectively. Results (1) The immunohistochemical results showed that IKKε was highly expressed in epithelial ovarian cancer specimens with the expression rate 66.1% (78/118), compared with normal ovarian tissue with the expression rate 35.0% (7/20), which exhibited statistically significant difference (χ2=6.993, P=0.008). The expression of IKKε protein was correlated with International Federation of Gynecology and Obstetrics (FIGO) stage, histological grade, the level of CA125 in preoperative serum and distribution of the tumor (P0.05). (2) IKKε was widely overexpressed in different levels in SKOV3, OV2008, C13, A2780S, A2780CP, OV4, OV5, OV8, and CAOV3 cells, and the expression of IKKε decreased as the increase of the concentration of IKKε inhibitor (0.1 and 0.5 μmol/L) in OV2008, C13, A2780S, and A2780CP cells after 48 hours treatment. Different concentrations of IKKε inhibitor (0.05, 0.1, 0.5, 1, 5, 10, and 25 μmol/L) significantly inhibited the proliferation of OV2008, C13, A2780S, A2780CP, and SKOV3 cells in a concentration-dependent manner (P<0.05), and the half maximal inhibitory concentration (IC50) was 0.43, 0.86, 0.10, 0.19, and 0.24 μmol/L, respectively. The cell apoptotic rate of OV2008, C13, A2780S, A2780CP, and SKOV3 cells was significantly increased after 48 hours treatment of IKKεinhibitor with the concentration of 0.1 and 0.5 μmol/L (P<0.05). Conclusions The IKKε protein in epithelial ovarian cancer specimens and cells is overexpressed. IKKε inhibitor could inhibit cellular proliferation and induce apoptosis in a concentration-dependent manner. Together, the result indicated that IKKε may be a candidate target for the treatment of ovarian cancer in future.
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Objective To detect and explore the expression and clinical significance of dual specificity tyrosine phosphorylation regulated kinase1b (Dyrk1b) in the specimens and cells of cervical lesions. Methods (1)All the data were collected from 75 patients with cervical cancer and 52 cases with squamous intraepithelial lesion(SIL)admitted in the First Affiliated Hospital of Dalian Medical College during Jan. 2011 to Dec. 2013 and confirmed by pathological examination, included 60 cases of stageⅠand 15 cases of stageⅡ, 12 cases with low-grade squamous intraepithelial lesion(LSIL)and 40 cases with high-grade squamous intraepithelial lesion(HSIL). While, 28 cases with chronic cervicitis were chosen as the control group. The protein expression of Dyrk1b was detected by immunohistochemistry among the four groups.(2)The expression of Dyrk1b in HeLa and SiHa cells were detected by western blot method and the expression of Dyrk1b protein were also detected after treatment of AZ191 (5, 10 μmol/L) for 48 hours in HeLa and SiHa cells.(3)The cellular survival and proliferation of HeLa and SiHa cells treated by different concentrations of AZ191(2.5, 5, 10, 25, 50, 100 μmol/L)for 48 hours were detected by methyl thiazolyl tetrazolium (MTT) assay.(4)The rate of apoptosis of HeLa and SiHa cells was detected by flowcytometry after treatment of AZ191 (5, 10μmol/L) for 48 hours. Results (1)The positive rates of Dyrk1b protein in chronic cervicitis, LSIL, HSIL and cervical squamous cancer by immunohistochemistry were 11%(3/28), 1/12, 42%(17/40)and 71%(53/75), respectively. The expression of Dyrk1b in cervical squamous cancer and HISL were higher than those in LSIL and chronic cervicitis (P0.05). Expression of Dyrk1b was correlated with stromal invasion depth of cervical cancer (P0.05). (2) Dyrk1b protein was expressed in different levels in HeLa and SiHa cells, and the expression of Dyrk1b was decreased gradually as the increased of the concentration of AZ191 in both HeLa and SiHa cells by treatment of AZ191 for 48 hours. (3) Different concentration of AZ191 treated on cervical cancer cells could inhibit the cellular proliferation and induce cell apoptosis in a concentration-dependent manner(P<0.01), concomitant to the decreased cell survival rate. The apoptosis rate of HeLa and SiHa were increased significantly after 10μmol/L AZ191-treatment for 48 hours, but no any difference induced by 5 μmol/L AZ191-treatment compared to control group. Also,there was no any difference between Hela and SiHa cells in either inhibitory effect or apoptosis rate induced by AZ191. Conclusions Dyrk1b is over-expressed in either specimens or cells of cervical cancer. The expression of Dyrk1b protein in cervical lesions is increased as the progression of disease. Dyrk1b inhibitor AZ191 could inhibit cellular proliferation and induce apoptosis in a concentration-dependent manner in cervical cancer cells.
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Objective To explore the safety and efficacy of small margin in nephron sparing surgery for early localized renal cell carcinoma (RCC). Methods A total of 325 cases of RCC with normal contralateral kidney and staged as Tla were retrospectivly studied.According to the margin size,125 cases were with surgical margin ≤ 5 mm (group ≤ 5 mm),102 cases with margin 6-9 mm (group 6-9 mm) and 98 cases with margin ＞ 10 mm (group ＞ 10 mm).The margin size and status was pathologically evaluated and clinical results including local recurrence,distant metastasis and overall survival rate were followed up and comparatively analyzed. Results None of the patients had positive surgical margins.The mean and median margin sizes were 2.2 and 2.0 mm for group ≤ 5 mm,6.7 and 6.0 mm for group 6-9 mm and 11.8 and 12.0 mm for group ＞ 10 mm.The difference was statistically significant (P=0.025).The mean and median follow-up time for all the patients were 79 and 83 months (range 15-132 months),with 69 and 73 months (range,15-130 months) for group ≤ 5 mm,83 and 86 (range,17-132 months) for group 6-9mm and 82 and 82 (range 60-103 months) for group ＞ 10 mm.Three patients in group ≤ 5 mm,5 in group 6-9 mm and 2 in group ＞ 10 mm died of no-cancer related disease during follow-up.One patient in group ≤ 5 mm (0.74％) experienced ectopic recurrence in the same kidney and one in group 6-9 mm was detected local recurrence in situ (0.98％).No distant metastasis was detected in all the patients.The overall 5-year survival rate for patients in groups ≤ 5mm,6-9 mm and ＞ 10 mm were 99.2％,99.0％ and 98.0％,respectively.(Kaplan-Meier survival analysis,Log Rank,x2 =1.511,P=0.470). Conclusions Small margin in nephron sparing surgery is safe and effective in treating RCC with stage T1a,which provides excellent renal function preservation,favorable long-term progression-free survival rate,and is not associated with an increased risk of local recurrence.
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Objective To investigate the expression and significance of caveolin-1 in breast carcinoma.Methods Using immunohistochemical method the protein expression of caveolin-1 were analyzed in 105 cases of breast carcinoma and 50 cases of non-cancerous breast tissues.The relationship between caveolin-1 expression and CK5/6,EGFR,and E-cadherin expression was investigated.Clinical data of 105 cases of breast carcinoma were retrospectively analyzed.Results Among 105 cases of breast carcinoma,there were 20 cases of basal-like subtype,22 cases of luminal subtype A,23 cases of luminal subtype B,23 cases of HER2 over-expressing subtype,17 cases of normal breast-like subtype.Positive rate of caveolin-1 was significantly lower in breast carcinoma than in non-cancerous breast tissues (24.8％ vs.88.0％,P ＜ 0.05).Positive rate of caveolin-1 (75.0％) was higher in breast carcinoma than in luminal subtype A ( 4.8％ ),luminal subtype B ( 17.4％ ),HER2 over-expressing subtype ( 17.4％ ) or normal breast-like subtype( 11.8％ ),all P ＜0.05.Caveolin-1 expression was associated with expression of CK5/6 and EGFR(P ＜0.01 ).In univariate analysis,positive caveolin-1 was associated with higher lymph node metastasis rate (18/26,69.2％ )than negative (37/79,46.8％ ),P =0.047,and shorter 5-year-disease-free survival (38.46％ vs.74.68％,P =0.0004 ),but in multivariate analysis caveolin-1 was not an independent predictor of 5-year- disease-free survival (P ＞ 0.05).Conclusions Caveolin-1 can be seen as a screening mark of basal-like breast carcinoma,it may promote the invasiveness of breast cancer cells,but it is not an independent prognostic predictor of breast cancer patients.
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Objective To investigate the expression of E-cadherin and matrix metallopeptidase 7(MMP-7)and their significance in basal-like and Her-2 over-expressing breast carcinoma. Methods The protein expression of E-cadherin and MMP-7 were analyzed by immunohistochemical method in 20 cases of basal-like breast carcinoma and 21 cases of Her-2 over-expressing breast carcinoma.Statistical method was used to analyze the clinicopathological performance.Patients were followed up from 6 to 60 months.Results The ratio of basal-like breast carcinoma and invasive ductal carcinoma was 3.4%(20/501).the mean age was 49 years.The mean size of basal-like breast carcinoma(2.1 cm)was smaller than that of Her2 over-expressing breast carcinoma(3.0 cm)(P<0.05).The 5-year survival rate of patients with basal-like breast carcinoma was obviously(40.0%) lower than that of Her-2 over-expressing breast carcinoma (71.4%)(P<0.05),though,the 3-year survival rate was similar between the two groups.The protein expression of E-cadherin was down-regulated and even deficient in basal-like breast carcinoma(P<0.05).The protein expression of MMP-7 was higher in basal-1ike breast carcinoma than that in Her-2 overexpressing breast carcinoma(P<0.05).Conclusion Down-regulated expression of E-cadherin and upregulated expression of MMP-7 may be one of the mechanisms leading to high metastasis of basal-like breast carcinoma.
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<p><b>OBJECTIVE</b>To investigate the incidence and associated factors of multicentricity in renal cell carcinoma (RCC) in Chinese patients.</p><p><b>METHODS</b>One hundred and two kidney samples from radical nephrectomy due to RCC were step sectioned at 3 mm intervals and examined. All tissue abnormalities were removed, stained and examined for multicentricity. Then, on each slice of the sample, both the parenchymal margin of 15 mm beyond the pseudocapsule and tissue around the renal sinus were continuously sectioned and examined for completeness of the pseudocapsule and vascular and lymph node invasion. The relationship between muliticentricity and other pathological parameters was evaluated.</p><p><b>RESULTS</b>The incidence of multicentricity was 15.7% (16/102); it was significantly lower in primary tumors < or = 4.0 cm than in tumors > 4.0 cm (4.9%, 2/41 vs 23.0%, 14/61; chi(2) = 6.055, P = 0.014). The incidence was 9.8% (8/82) in tumors without vascular invasion and 40.0% (8/20) in those with it (P = 0.003, Fisher's exact test). The incidence of multicentricity was 1.9% (1/53) in tumors with a complete pseudocapsule and 30.6% (15/49) in those without it (chi(2) = 15.885, P = 0.000). The grade, stage, subtypes and lymph node invasion of the primary tumor were not significantly associated with multicentricity. Multiple logistic regression analysis showed that pseudocapsular incompleteness and vascular invasion were two significant predictors of RCC multicentricity (P = 0.005 and 0.023).</p><p><b>CONCLUSIONS</b>The incidence of multicentricity of RCC in this group of patients was in accordance with published studies. Multifocality was significantly associated with tumor size, pseudocapsule completeness and vascular invasion. NSS should be limited to tumors less than 4.0 cm when the contralateral kidney is normal and careful long-term follow-up is necessary in tumors with positive vascular invasion and incomplete pseudocapsule.</p>
Subject(s)
Aged , Female , Humans , Male , Middle Aged , Carcinoma, Renal Cell , Epidemiology , Pathology , General Surgery , China , Epidemiology , Incidence , Kidney Neoplasms , Epidemiology , Pathology , General Surgery , Lymphatic Metastasis , Neoplasm InvasivenessABSTRACT
Objective To study the incidence of multi ce ntricity in renal cell carcinoma (RCC). Methods 102 kidn eys from radical nephrectomy for RCC were step sectioned at 3 mm intervals and a ny abnormal tissue were removed, stained and examined. The tissue 2.0 cm beyond pseudocapsule and tissue of renal hilum were continueously or interruptedly sect ioned and examined for any tumor invasion of the pseudocapsule, micro-multifoca l carcinoma and vascular invasion beyond pseudocapsule. The relationship between the pathological findings of the primary tumors and multicentricity was evaluat ed. Results The total incidence of multifocal carcinoma in this group was 15.7%.It was 4.9% in primary tumors 4 cm or less and 23.0% in tumors larger than 4 cm (P= 0.014). Vascular invasion and interrupted pseud ocapsule were two significant predictors of multicentricity of RCC (P=0.017 and 0.006). Conclusions In the RCC patients with normal contralateral kidneys, nephron-sparing surgery should be performed only in tumo rs 4 cm or less. In patients nephron-sparing surgery is imperative, even tumors 7 cm or less might be in consideration, but intensive followup is necessary due to the increased incidence of multicentricity.In most cases, a partial nephrect omy instead of tumor enucleation should be performed.
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0.4).ConclusionsThese data denote that biological behaviors and malignant features of multifocal tumors are very similar to those of primary ones.In addition,considering the significant relationship of multicentricity with vascular invasion and incompleteness of pseudocapsule of RCC,it is suggested that the secondary tumors might be more likely the result of intrarenal metastasis of the primary tumor rather than the results of multifocal genesis.
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Objective To assess the necessary excision of normal parenchyma tissue in nephron sparing surgery for renal cell carcinoma. Methods 102 specimens from radical nephrectomy for RCC were step sectioned at 3 mm intervals and examined.The tumor and field 20 mm beyond pseudocapsule were continuously sectioned and examined for completeness of pseudocapsule,the presence of micro multifocal carcinoma and for vascular and parenchyma invasion beyond pseudocapsule. Results 49 (48.0%) of the 102 specimens were void of intact pseudocapsule.The presence of extra pseudocapsule cancerous lesions was within 0～8 mm [(1.2?1.9) mm] beyone the capsule,with 30.4% of them within the field of 1～8 mm.with the statistic method of one side analysis of frequency,the percentile value of P97.5 and P100.0 was 7.4 and 8.0 mm respectively. Conclusions These data denote that when nephron sparing surgery is done for renal cell carcinoma,at least 10 mm of normal parenchyma tissue beyond the pseudocapsule should be excised with the tumor.The nucleation techniquc should not be encouraged.