The clinical significance and underlying correlation of pStat-3 and integrin αvß6 expression in gallbladder cancer.

BACKGROUND: Both phosphorylated signal transducer and activator of transcription 3(pStat-3) and integrin αvß6 can play vital role in the development and progression of cancer. However, little is known about their expression correlation and clinical significance in gallbladder cancer(GBC). OBJECTIVE: The aim of our present study was to investigate the expression of pStat-3 and integrin αvß6, two proteins' correlation and their clinical significance in GBC tissues. RESULTS: The expression of pStat-3 and integrin αvß6 were both significantly associated with T stage, lymph node metastasis status, TNM stage (P=0.008, P=0.000, P=0.000 and P=0.036, P=0.001,P=0.000,respectively). IHC and Western blot showed their expressions in GBC tissues were higher than that in paraneoplastic tissues. Moderate positive correlation existed between the two proteins (r =0.349, P <0.001). The survival analysis by Kaplan-Meier and Cox regression model showed that GBC patients with pStat-3 or integrin αvß6 positive expression had a significantly poorer 2-year survival rate (P = 0.002 and 0.000, the log-rank test, respectively), and either marker could act as unfavorable independent prognostic factors(RR=1.907, P=0.021 and RR=2.046, P=0.038). MATERIALS AND METHODS: The expression levels of pStat-3 and integrin αvß6 were analyzed in GBC cancerous and paraneoplastic tissues of 97 cases via immunohistochemistry(IHC) and further validated by western blot method. Besides, SPSS software was used to observe their clinical significance as well as the two proteins' correlation. CONCLUSION: pStat-3 and integrin αvß6 were indicators of tumor's progression and poor prognosis of patients with GBC. And the further study involving them may provide a helpful therapeutic target in prevention and treatment of GBC patients.

Integrin ß6 can be translationally regulated by eukaryotic initiation factor 4E: Contributing to colonic tumor malignancy.

It is well known that both eukaryotic initiation factor 4E (eIF4E) and integrin αvß6 can contribute to malignant behavior of colon cancer. We have found that integrin αvß6 and eIF4E were co-expressed and positively correlated in colon cancer tissues. Recently, deregulation of the protein synthesis apparatus has begun to gain attention as a major participant in cancer development and progression. However, the regulation of integrin ß6 expression at translational level has never been investigated before. In present study, gene-silencing technique for eIF4E by small interfering RNA (siRNA) was used in all the subsequent experiments, in order to investigate whether eIF4E could translationally regulate expression of integrin ß6 in colon cancer SW480 and HT-29 cell lines. Additionally, the subsequent effects of eIF4E knockdown on cellular malignant behavior were observed. siRNA in SW480 and HT-29 transfectants. Subsequently, protein expression of ß6 was markedly suppressed, while mRNA expression of ß6 showed no significant variation before and after eIF4E RNA interfering. Therefore, it could be seen that eIF4E could upregulate the expression of ß6, without effect on ß6 mRNA expression. More importantly, after treated with eIF4E siRNA, cellular migratory capacity on fibronectin of HT-29 and ß6-transfected SW480 as well as their survival to 5-FU was decreased distinctly. Expression of integrin ß6 could be translationally regulated by eIF4E, which subsequently contributed to tumor malignancy through enhancing cellular migration, survival, anti-apoptosis, and chemoresistance of colon cancer in vitro. Thus, targeting eIF4E in integrin αvß6 expressing tumors can be a potential therapeutic strategy for patients with colon cancer.

Tolerance of gastric cancer cells to 5-fluorouracil mediated by integrin αvβ6 through extracellular signal-regulated protein kinase / 中华消化外科杂志

Objective To explore the role of integrin αvβ6 in mediating the tolerance of gastric cancer AGS cells to 5-fluorouracil, and to determine whether direct β6-extracellular signal-regulated protein kinase(ERK) binding is involved. Methods Gastric cancer AGS cells in the logarithmic phase were incubated with either 5-fluorouracil for 24 hours ( control group), with 0.1 g/L of mouse anti-αvβ6 monoclonal antibody 10D5 for 6 hours and then with 5-fluorouracil for24 hours ( 10D5 group), with IgG2a and 5-fluorouracil ( IgG2a group), or with 5-fluorouracil and 20 μnol/L of ERK inhibitor PD98059 for 24 hours ( PD98059 group). Cell proliferation,apoptosis and the expression of Bcl-2 and caspase-3 protein were detected by MTT assay, flow cytometry and western blotting, respectively. All data were analysed by one-way analysis of variance and LSD test. Results The cell inhibition rates of the control group, 10D5 group, IgG2a group and PD98059 group were 28.1% ±2.7%,84.5% ± 1.6%, 31.4% ±5.2%, 86.7% ±5.2%, respectively, with a significant difference ( F = 342. 5, P ＜0.05). The apoptosis rates of the control group, 10D5 group, IgG2a group, and PD98059 group were 6.6% ±1.4%, 30.6% ± 2.4%, 8.1% ± 1.3%, 36.0% ±4.0%, respectively, with a significant difference among the four groups (F = 105.4, P ＜0.05 ). There was a significant difference in the expression of caspase-3 and Bcl-2 among the four groups (F=292.1, 181.6, P＜0.05). Conclusion Integrin αvβ6 can mediate the tolerance of gastric cancer AGS cells to 5-fluorouracil through direct β6-ERK binding.

The role of integrin ανβ6 in regulation of extracellular matrix degradation of colon cancer cells / 中华老年医学杂志

Objective To investigate the roles of integrin ανβ6 in the invasion of colon cancer cells and its molecular mechanisms for regulation of the extracellular matrix (ECM) degradation.Methods The RNA interfering technique was used to downregulate the expression of ανβ6 in colon cancer cells. The expression of ανβ6 in transfected cells was determined by RT-PCR and Western blot.The cell invasive capacity was evaluated by reconstituted basement membrane invasion assay. Western blot and gelatin zymography were used to determine whether the reduced αvβ6 expression affects extracellular signal-regulated kinase (ERK), P-ERK, urokinase-type plasminogen activator (uPA) and matrix metalloproteinases (MMPs) expressions. [3H]-labelled type Ⅳ collagen degradation assay was performed to asess if supression of integrin ανβ6 inhibits extracellular matrix degradation.Results Specific siRNA inhibited the expression of ανβ6 in both the protein and mRNA levels in HT29 cells. Down regulation of integrin ανβ6 inhibited ERK, P-ERK1/2 expressions, and the secretion of uPA. pro-MMP-9 and pro-MMP-2 in tumor conditioned medium. Supression of integrin ανβ6 inhibited MAPK dependent [3H]-labelled type Ⅳ collagen degradation. Conclusions These data in our study demonstrats that integrin ανβ6 plays important roles in the invasion of colon cancer cells. The ανβ6 regulates the secretion levels of uPA, pro-MMP-9, pro-MMP-2 and the ECM degradation through the MAPK pathway.

A new method of NOTES:experimental study of totally transtracheal endoscopic thyroidectomy on animals / 中国现代普通外科进展

Objective:To investigate the feasibility and safety of a new method of natural orifice transluminal endoscopic surgery(NOTES) -totally transtracheal endoscopic thyroidectomy(TTET) .Methods:Three miniature swines and 6 beagle dogs were underwent TTET.Under general anesthesia,special designed endotracheal tube with 2-channel was used and endoscope and instruments were inserted through the respective channel.Incision of tracheal anterior wall was accomplished and partial or subtotal thyroidectomy was performed.Finally,the defects in the trachea were sutured with ENDO STITCH instrument.Results:Partial thyroidectomy was successfully accomplished on 3 pigs and subtotal thyroidectomy was done on 6 dogs.No serious complications such as anoxia,asphyxia,airway obstruction and death occurred during the operation.Animals were sacrificed 2h after the procedure and incision of trachea was found to be closely sutured.There were no subcutaneous emphysema and haematoma formation.Conclusion:Preliminary experimental results showed the feasibility and safety of TTET.Transtracheal access maintains the integrity of cervical tissues and achieves an optimal cosmetic outcome.TTET may open up a new field of NOTES on thyroid surgery.

Apoptosis of acitretin-induced cutaneous squamous cell carcinoma cells and effects on cuspases expressions / 中国综合临床

Objective To investigate the inducing effect of acitrotin on the growth and apoptosis of human cutaneous squamous cell carcinoma cell line SCC13 and on caspases expression.Methods Human cutaneous squa-mous cell carcinoma cell line SCC13 was treated with five different concentrations of acitrefin [5×10-7,1×10-6,5×10-6,1×10-5,5×10-5mol/L].Cell proliferation was evaluated by MTT assay.Apoptosis was assessed by en-zyme-linked immunosorbent assay.The cell cycle was assessed by flow cytometry.The protein expressions of caspase-8 and caspase-9 were examined with Western blot.Results Acitretin inhibited the growth ( F = 83.64,96.34 and 123.17, respectively on the first, third and fifth day)and induced the apoptosis of SCC13 cells(F=74.45,107.37,and 64.28, respectively on the first, third and fifth day) in a dose- and time-dependent manner(P<0.05).Acitretin altered cell cycle distribution of SCC13 cells as compared with controls, the G1-phase population increased by 77.66% 72 hours after acitretin treatment, while the control increased only by 63.55%.An active fragment of caspase-8 occurred following 12 hours treatment of acitretin on SCC13 cells, whereas the caspase-9 active fragment occurred 24 hours after acitretin treatment, which increased time-dependently (P<0.01).Conclusion Acitretin plays an important role on the growth inhibition and apoptosis of cutaneous squamous cell carcinoma cells, which may be affected through both Fas receptor way and mitochondria way.

Role of integrin ?v?6 in proliferation and apoptosis of gastric cancer AGS cells / 中国现代普通外科进展

Objective: To explore the role of integrin ?v?6 in proliferation and apoptosis of gastric cancer AGS cells. Methods: Gastric cancer AGS cells were treated with ?v?6 monoclonal antibody 10D5 or the negative control mouse immunogloblins IgG2a. Cell viability was measured by MTT assay, cell apoptosis was detected by FCM, and caspase-3 activity was examined by Western blot. Results: Compared with the control and IgG2a group, the apoptotic rate and caspase-3 activity of AGS cells treated with 10D5 increased, meanwhile cells survival rate decreased. There were significant differences of the indexes between the 10D5 group and the other two groups (P0.05). Conclusion: Integrin ?v?6 plays an important role in the proliferation and apoptosis of gastric cancer AGS cells.