ABSTRACT
Objective To explore the expression of cullin 4A (CUL4A) in cholangiocarcinoma tissues and its clinical significance.Methods Primary fresh cholangiocarcinoma tissues (n =35) and normal bile duct tissues (n =15) from patients who underwent curative surgery in Binzhou People's Hospital of Shandong Province from February 2011 to December 2013 were collected.The expressions of CUL4A mRNA were detected by quantitative real-time PCR (qRT-PCR).Then,cholangiocarcinoma tissues (n =72) and normal bile duct tissues (n =36) from patients who underwent curative surgery in Binzhou People's Hospital of Shandong Province from January 2008 to January 2014 were collected.The expressions of CUL4A protein were tested by immunohistochemistry.The relationships between the expression of CUL4A and the patients' clinicopathologic features and prognosis were analyzed.Results The expression of CUL4A mRNA in cholangiocarcinoma tissues was obviously higher than that in normal bile duct tissues (3.876 ±0.975 vs.1.216 ±0.265),and the difference was statistically significant (t =12.23,P ＜ 0.001).The positive rates of CUL4A protein in cholangiocarcinoma and normal bile duct tissues were 70.8％ (51/72) and 2.8％ (1/36) respectively,and the difference was statistically significant (x2 =44.524,P ＜ 0.001).The expression of CUL4A protein in cholangiocarcinoma was related to the differentiated degree (x2 =4.341,P =0.037),neural invasion (x2 =8.326,P =0.004),TNM stage (x2 =7.745,P =0.005),lymph node metastasis (x2 =3.869,P =0.049) and vascular invasion (x2 =5.555,P =0.018).Survival analysis results showed that the median survival time of CUL4A positive patients was significantly shorter than that of CUL4A negative patients (32.40 months vs.51.30 months),and the difference was statistically significant (x2 =6.561,P =0.011).Conclusion The expression of CUL4A in cholangiocarcinoma tissues is higher than that in normal bile tissues.CUL4A plays an important role in the process of tumor invasion and metastasis and indicates poor prognosis,which may become a potential target for the diagnosis and therapy of the cholangiocarcinoma.
ABSTRACT
Objective To investigate the expressions of TRIM59,Twist and E-cadherin in hepatocellular carcinoma and their clinical significance. Methods The expressions of TRIM59,Twist and E-cadherin protein were tested by immunohistochemistry in 80 cases of hepatocellular carcinomas and the adjacent paracancerous tissues. Results The positive rate of TRIM59 in hepatocellular carcinoma was significantly higher than that in the adjacent paracancerous tissue(76.3%vs. 8.0%,P<0.05). Significant difference was also observed in the expres-sion rate of Twist between the hepatocellular carcinomas and the paracancerous tissue(66.3%vs. 6.0%,P<0.05). The positive rate of E-cadherin in hepatocellular carcinoma was significantly lower than that in the adjacent para-cancerous tissue(27.5%vs. 90.0%,P<0.05). The differences of the expression of TRIM59 in hepatocellular carci-noma of pathological grading,tumor differentiation,vascular invasion and clinical TNM stage were significant(P<0.05). The differences of the expression of Twist in hepatocellular carcinoma of pathological grading ,differentia-tion,vascular invasion and clinical TNM stage was also significant(P<0.05,respectively). The differences of the expression of E-cadherin in hepatocellular carcinoma of pathological grading,differentiation,vascular invasion and clinical TNM stage were also significant(P<0.05,respectively). Significantly positive correlation was also found between TRIM59 and Twist by using spearman correlation analysis(P<0.05). Negative correlations were observed between TRIM59 and E-cadherin(P < 0.05),and between Twist and E-cadherin(P < 0.05). Survival analysis showed that TRIM59 expression was an independent prognostic factor in hepatocellular carcinoma. Conclusion TRIM59,Twist and E-cadherin protein expression might be associated with the development,invasion,and metas-tasis of hepatocellular carcinoma. TRIM59 may become a new target gene for the treatment of human hepatocellular carcinoma.
ABSTRACT
Objective To investigate the influence of proline-rich tyrosine kinase 2 (Pyk2)gene RNA interference on proliferation,invasion and migration of Hep3B hepatocellular carcinoma cells.Methods The Pyk2 gene RNA interference vector was transfected in Hep3B hepatocellular carcinoma cells by lipofectamine. The Hep3B cells divided into three groups:siRNA group (the vector with Pyk2 RNAi gene was transfected), negative control group (the vector without Pyk2 RNAi gene was transfected),and blank control group (no vectors was transfected).Pyk2 mRNA and protein were detected using reverse transcription reverse transcription-poly-merase chain reaction (RT-PCR)and Western blotting.The biological behavior including cell proliferation,inva-sion and migration were detected by 3-(4,5-dimethyl-2-thiazoly)-2,5-diphenyl-2H-tetrazolium bromide (MTT), transwell and wound healing assay,respectively.Results The expression of Pyk2 mRNA of Hep3B cell line in siRNA group (0.1 6 ±0.03)was significantly decreased than those in negative group (0.74 ±0.1 3)and blank control group (0.77 ±0.1 6),with statistically significant differences (t=51 .46,P=0.000;t=53.21 ,P=0.000).The expression of Pyk2 protein of Hep3B cell line in siRNA group (0.24 ±0.06)was significantly decreased than those in negative group (0.83 ±0.05)and blank control group (0.91 ±0.06),with statisti-cally significant differences (t=57.29,P=0.000;t=68.53,P=0.000).The cell proliferation inhibition rate at 48 hours in siRNA group (26.1 7%±0.28%)was significantly raised than those in negative group (9.28%± 0.22%)and blank control group (6.47%±0.31%),with statistically significant differences (t=31 .45,P=0.004;t=34.64,P=0.002).The number of transmembrane cells in siRNA group (32.5 ±8.5)/1 0 HP was significantly declined than those in negative group (98.4 ±1 2.3 )/1 0 HP and blank control group (1 1 2.6 ± 1 1 .3)/1 0 HP,with statistically significant differences (t=95.64,P=0.000;t=1 05.1 7,P=0.000).The wound healing assay in siRNA group (28.1 7%±1 .46%)was significantly lower than those in negative group (77.38%±2.24%)and blank control group (79.41%±3.1 7%),with statistically significant (t=85.86,P=0.000;t=89.37,P=0.000).Conclusion Pyk2 gene involves the proliferation,invasion and migration of Hep3B cells,which has close correction with development and metastasis of hepatocellular carcinoma.Pyk2 gene is very helpful to become a molecular target for the diagnosis and treatment of hepatocellular carcinoma.
ABSTRACT
Objective To investigate the effects of MMI-166 on apoptosis and apoptosis-related protein expression of human pancreatic cancer SW1990 cells and its transplanted tumor,and explore possible mechanism.Methods The human pancreatic cancer xenograft model was constructed by using human pancreatic cancer SW1990 cells.Tumor-bearing nude mice were randomly divided into control and MMI-166 groups,and they were treated with normal saline or MMI-166 (200 mg · kg-1 · d-1) for 28 days.Apoptosis index (AI),p53,c-Myc,Bax,Bcl-2,Survivin,Caspase-1,Fas proteins were detected by deoxynucl-eotidyl transferase-mediated nick end labeling (TUNEL method) and Western blot.MMI-166 of different concentrations (0,50,100 μg/ml) were used to treat human pancreatic cancer SW1990 cell for 24 h.The c-Myc,Survivin proteins expressions were measured by Western blotting.Results Apoptosis index in MMI-166 group was 81.1 ±7.9,which was significantly higher than that in control group (21.3 ±2.2,P =0.000) ; the expressions of c-Myc,Survivin were 7715 ± 2229,4594 ± 1240,which were significantly higher than those in control group (16870 ± 2446,15208 ± 1903,P =0.000) ; the expressions of p53,Bax,Bcl-2,Caspase-1,Fas were not significantly different from those in control group.After 50,100 μg/ml MMI-166 treatment,the expression of c-Myc was significantly down-regulated (0.098 ± 0.003,0.073 ± 0.008 vs.0.169 ± 0.007,F =189.361,P ＜ 0.05) ; and the expression of Survivin was not significantly changed.Conclusions MMI-166 may induce cell apoptosis of SW1990 by down-regulating the expression of c-Myc.
ABSTRACT
ObjectiveTo study the expressions of Survivin and Bcl-2 in carcinoma of the ampulla of Vater and their clinical significance.Methods The expressions of Survivin and Bcl-2 proteins were tested by EnVision immunohistochemistry in 40 cases of ampullary carcinomas,and 8 cases of normal ampulla of rater as control.ResultsThe positive expression of Survivin and Bcl-2 in the tissues from the ampullary carcinoma was significantly higher than that in the normal controls(82.5％,72.5％ vs 0％,12.5％,P ＜0.05).The expression of Survivin and Bcl-2 had no relations with the sex,age,tumor size,histological types,and clinical stages( P ＞ 0.05 ).The differences of the expression of Survivin and Bcl-2 in ampullary carcinoma of duodenal invasion,pancreatic invasion,and lymph node metastasis were significant(P ＜ 0.05) ; Significantly positive correlation was found between the expression of Survivin and Bcl-2 by using spearman correlation analysis( r =0.647,P ＜ 0.05 ).Conclusions Survivin and Bcl-2 may play an important role in tumorigenesis of ampullary carcinoma.Combined detection of Survivin and Bcl-2 may be useful to determine the degree of malignancy and the progress of the ampullary carcinoma.
ABSTRACT
Objective To investigate the differentiated thyroid carcinoma diagnosis and treatment options. Methods From Feb. 2002 to Jan. 2008,78 patients received different surgical resection regarding the type of tumor size,number of tumor,ages and jugular lymphatic metastasis. Patients with unilateral differentiated thyroid carcinoma underwent the resection of ipsilateral isthmus of thyroid lobe or plus partial contralateral gland,and those with bilateral-lobe underwent total thyroidectomy or near-total thyroidectomy. High-risk patients (age >45 years,tumor size >4 cm,tumor size ≤4 cm,but surpass the envelop of thyroid) were performed by functional neck dissection or lymph node dissection of central region (Ⅵ area) besides postoperative endocrine therapy. Results Eleven cases underwent the resection of ipsilateral lobe with isthmus, 19 cases underwent surgical removal of ipsilateral lobe with isthmus plus partial contralateral gland,26 cases underwent near-total thyroidectomy and 22 total thyroidectomy. 25 cases underwent functional neck dissection, 23 cases underwent neck dissection of central region. There were 68 papillary thyroid carcinoma (87. 18%), 10 follicular thyroid carcinoma (12. 82%). There were 26 cases with lymphatic metastasis of Ⅵ area. Postoperative complications included 12 cases (15.38%) with deadlimb caused by hypocalcemia, 8 cases (10.26%) with transient recurrent nerve paralysis,2 cases (2.56%) with permanent injury of recurrent laryngeal nerves, 3 cases (3. 58%) with chylous fistula. Seventy-four(94. 87%) cases were followed up postoperatively for a period from 6 months to 6 years,which showed that no death occurred,but 6 relapsed with jugular lymphatic metastasis,after reoperation no distant metastasis occurred. Survival rate was 97. 30% (72/74). Conclusion Treatment of the differentiated thyroid carcinoma should be based on the size of tumor,number of tumor,age and jugular lymphatic metastasis. Lymph node dissection of central region was necessary for high-risk patients of differentiated thyroid carcinoma.