OBJECTIVETo explore the role of S100A4 in the epithelial-mesenchymal transition (EMT) in esophageal squamous cell carcinoma and its possible molecular mechanism.

METHODSThree chemically synthesized S100A4 siRNA sequences were transiently transfected into esophageal carcinoma EC9706 cells. EC9706 cells transfected with negative siRNA, lipofectamine 2000, and vacant EC9706 cells were used as control. Fluorescence quantitative RT-PCR and Western blot were used to detect the inhibition rate of S100A4 siRNA. S100A4 siRNA2 with the best inhibition rate was chosen to transiently transfect into EC9706 cells under the same conditions. The EC9706 cells transfected with negative siRNA, lipofectamine 2000 and vacant EC9706 cells were also used as control. Fluorescence quantitative RT-PCR and Western blot were used to detect the mRNA and protein expressions of E-cadherin, vimentin and snail. The morphology of EC9706 cells was observed under an inverted microscope. Boyden chamber and scratch test were used to detect the invasion and migration ability of EC9706 cells, and CCK8 assay was used to detect the proliferation ability of EC9706 cells. EC9706 cells transfected with S100A4 siRNA2 were further transfected with snail eukaryotic expression vector. The EC9706 cells transfected with S100A4 siRNA, EC9706 cells transfected with snail eukaryotic expression vector and vacant EC9706 cells were used as control. The above indexes of all the groups were observed, too.

RESULTSThe S100A4 mRNA and protein expression levels of the S100A4 siRNA2 group were 0.417 ± 0.041 and 0.337 ± 0.039, the transmembrane cell number was 61.608 ± 8.937, the scratch healing distance was (0.216 ± 0.064) mm, the A value was 0.623 ± 0.084, the E-cadherin mRNA and protein levels were 0.619 ± 0.032 and 0.495 ± 0.034, the vimentin mRNA and protein levels were 0.514 ± 0.032 and 0.427 ± 0.028, the snail mRNA and protein levels were 0.573 ± 0.029 and 0.429 ± 0.041. These data were significantly different with the liposome group, the negative control group and the blank group (P < 0.05 for all). After the S100A4 siRNA2 treatment for 24 h, the appearance of EC9706 cells changed to epithelial cell morphology. The transmembrane cell number and the scratch healing distance of the S100A4 siRNA2+snail eukaryotic expression vector group were (69.382 ± 9.666) cells and (0.274 ± 0.029) mm, the A value was 0.823 ± 0.101, the snail mRNA and protein levels were 0.704 ± 0.037 and 0.625 ± 0.031, the vimentin mRNA and protein levels were 0.712 ± 0.046 and 0.609 ± 0.038, and these data were significantly higher than those of the Sl00A4 siRNA2 group (P < 0.05 for all). The E-cadherin mRNA and protein levels of the S100A4 siRNA2+eukaryotic expression vector group were 0.437 ± 0.038 and 0.381 ± 0.031, significantly lower than those of the S100A4 siRNA2 group (P < 0.05 for all). However, snail had no effect on the morphology of EC9706 cells.

CONCLUSIONSS100A4 may be involved in the EMT process of esophageal squamous-cell carcinoma by regulating the expression of snail and then plays a role in the invasion and metastasis of esophageal carcinoma.

Cadherins ; analysis ; Carcinoma, Squamous Cell ; metabolism ; pathology ; physiopathology ; Cell Line, Tumor ; Epithelial Cells ; Epithelial-Mesenchymal Transition ; Esophageal Neoplasms ; metabolism ; pathology ; physiopathology ; Humans ; Indicators and Reagents ; Lipids ; RNA, Messenger ; analysis ; RNA, Small Interfering ; analysis ; physiology ; S100 Calcium-Binding Protein A4 ; S100 Proteins ; antagonists & inhibitors ; genetics ; physiology ; Snail Family Transcription Factors ; Transcription Factors ; analysis ; genetics ; Transfection ; Vimentin ; analysis ; genetics

OBJECTIVE:To evaluate therapeutic efficacy of strontium dichloride (89SrCl2 ) combined with local radiotherapy for pain caused by malignant tumor bone metastasis. METHODS:115 patients with malignant tumor complicated with bone metasta-sis were analyzed retrospectively,and they received therapy regimen of 89SrCl2 combined with local radiotherapy. The amount of 89SrCl2 was calculated according to body weight of patients,and given by intravenous injection manner at the beginning of treat-ment,and then radiotherapy target section was confirmed according to iconography for irradiation. Effect of bone pain control,side effects (myelosuppression) and the effect of treatment on quality of life were evaluated at the end of treatment. RESULTS:After one month of treatment,there were 32 cases of complete remission(CR)and 62 cases of partial remission(PR),and total effec-tive rate was 81.7%;after one month of treatment,6 cases(5.1%)suffered from Ⅲ-Ⅳ degree myelosuppression;but there was none of Ⅲ-Ⅳdegree myelosuppression case after 2 months of treatment. The improvement of overall quality of life was not signifi-cant after treatment. Pain symptoms,general situation and sleep quality all improved significantly. CONCLUSIONS:89SrCl2 com-bined with local radiotherapy can control pain caused by malignant tumor bone metastasis and induce side effect,and improve quali-ty of life to certain extent.

Objective To study the inhibitory effect of arsenic trioxide(AS2O3)on the tumor growth of breast cancer line MDA-MB-435s implanted subcutaneously in nude mice and its mechanism.Methods BALB/C-nu/nu nude mice were subcutaneously injected with MDA-MB-435s breast cancer cells that are ER-positive,and treated with intraperitoneal injection of AS2O3 and DDP in different concentrations.The implanted tumor was weighed,and tumor inhibition rates were calculated.The expression of PTEN and Caspase-7 induced by AS2 03 were examined by immunohistochemical method.Results The growth of implanted tumor was markedly inhibited with DDP,low dose and high dose AS2O3,and the inhibitory rates were 48.68%,32.80%,66.67%respectively.The immunohisto chemical staining showed that the number of PTEN and Caspase-7 protein increased markedly(P＜0.05).Conclu sions AS2O3 inhibits the growth of human breast cancer cell implanted tumor.The molecular mechanism of AS2O3 on induction of apoptosis of breast cancer cells may be throush increasing the expression of PrrEN and Caspase-7(P ＜0.05).

Objective To observe the relationship between expression of retinoic acid receptor-β (RAR-β) in esophageal squamous cell carcinoma (ESCC) and chemotherapy response. Methods Fifty-two cases advanced ESCC patients treated by DDP and 5-FU, DDP 80 mg/m2, divided into 5 days;5-FU 375 mg/m2, dl-5. Immunohistochemistry was used to exmine the expression of RAR-β in ESCC. Fifty cases normal esophageal tissue were used as controls. Results RAR-β immunoreactivity was recognizd in both cytoplasm and nucleus, RAR-β positive rate was lower in ESCC compared with normal tissue (61.5%vs 92% ,P ＜0. 05 ). The 52 cases ESCC patients were treated 228 chemotherapy cycles, the overall response rate (OR) was 71.2%. The OR in RAR-β positive patients was 84. 4% (27/32), significant higher than RAR-β negative patients 50. 0% ( 10/20 ) ( P ＜ 0. 05 ). The time-to-progression ( TTP ) for RAR-β positive patients was 5.9 months, the median survival period was 12. 1 months, 2 years survival rate was 56. 7%;whereas TTP for RAR-β negative patients was 2. 1 months, the median survival period was 5.8 months,2 years survival rate was 32. 9%. There was signifcant difference between the 2 groups ( P ＜ 0. 05 ) .Conclusion RAR-β protein expression by immunohistochemistry may be a useful indicator to predict the chemotherapy response and clinical outcome for ESCC, meanwhile it may be an avenue for target therapy.

Objective:To investigate the effects of hematopoietic progenitor kinase 1 (HPK1) overexpression by construction of lentiviral vector on the proliferation and apoptosis of breast cancer MCF-7 and MDA-MB-231 cells,and to elucidate its possible mechanism.Methods:The cells were infected with the lentivirus overxpressing HPK1,and the MCF-7-HPK1 and MDA-MB-231-HPK1 cell lines were stably expressed HPK1;each cell line was divided into three experimental groups:blank group (untreated),control group (empty vector) and HPK1-overexpression group.The expression levels of HPK1 mRNA and protein in breast cancer cells in each group were detected by RTPCR and Western blotting methods,respectively.The cell proliferation rate was detected by MTT assay.The cell cycle and apoptotic rate were detected by flow cytometry.Transwell assay was used to analyze the cell migration ability.Western blotting method was used to measure the expression levels of caspase 3,PTEN,MMP-9,MMP-2,Ki-67and HPK1 proteins.Results:Compared with blank groups and control groups,the expression levels of HPK1 mRNA and protein in the both cell lines in HPK1 overexpression groups were significantly up-regulated (P＜0.05),the proliferation rates were significantly decreased (P＜0.05) and the apoptotic rates were significantly increased (P＜0.05),the number of cells crossing matrigel was significantly reduced (P＜0.05),the cell cycle of MCF-7 was blocked in G1 phase (P＜0.05),the expression levels of caspase 3 and PTEN proteins in HPK1 overexpression group were significantly increased (P＜0.05),and the expression levels of MMP-2 and MMP-9 proteins were significantly decreased (P＜0.05).Conclusion:HPK1 overexpression can inhibit the proliferation and migration of MCF-7 and MDA-MB-231 cells and induce apoptosis,which may be related to the up-regulation of caspase 3 and PTEN and down-regulation of MMP-9,MMP-2 and Ki-67.

BACKGROUNDAltered immunoresponse is associated with tumorigenesis and cancer progression. This study assessed the levels of tumor-infiltrating CD3 + or CD8 + T lymphocytes and interleukin-2 (IL-2) protein in radically resected non-small cell lung cancer (NSCLC) tissues to predict overall survival (OS) of the patients.

METHODSParaffin-embedded tissue specimens from 129 NSCLC patients were retrospectively collected for immunostaining of CD8 + , CD3 + , and IL-2 expression. Clinicopathological and survival data were collected and analyzed using the Chi-squared test, Kaplan-Meier curves, and the log-rank test or the Cox regression model.

RESULTSThe data showed a significant inverse association between CD8 + T lymphocyte levels and IL-2 expression (r = -0.927; P = 0.000) and between the levels of CD8 + and CD3 + T lymphocytes (r = -0.722; P = 0.000), but a positive association between CD3 + T lymphocyte levels and IL-2 expression (r = 0.781; P = 0.000) in NSCLC tissues. Furthermore, the levels of CD3 + and CD8 + T lymphocytes and IL-2 expression were associated with tumor stage (P = 0.023, 0.006, and 0.031, respectively) and the level of CD8 + T lymphocytes was associated with the patient gender (P = 0.024). In addition, the levels of CD8 + T lymphocytes were associated with an unfavorable 5-year OS, whereas patients with high levels of CD3 + T lymphocytes in tumor lesions and IL-2-expressing tumors had significantly better 5-year OS rates than patients with low levels.

CONCLUSIONSThe levels of CD8 + T cells in tumor lesions and IL-2 expression were both independent predictors of OS for these NSCLC patients. Thus, the detection of tumor-infiltrating CD3 + or CD8 + T lymphocytes and IL-2 expression could be useful to predict the prognosis of radically resected NSCLC patients.

CD3 Complex ; metabolism ; CD8-Positive T-Lymphocytes ; metabolism ; Female ; Humans ; Immunohistochemistry ; Interleukin-2 ; metabolism ; Lung Neoplasms ; immunology ; metabolism ; Lymphocytes, Tumor-Infiltrating ; metabolism ; Male ; Prognosis

Objective:To investigate the expression of transmembrane 4 super family 1 (TM4SF1)in breast cancer tissue,and to elucidate its clinical significance and explore the related molecular biological mechanisms. Methods:A total of 190 cases of human breast cancer,110 cases of paracancerous tissue and 110 cases of normal breast tissue were collected.Immunohistochemistry was used to detect the expression levels of TM4SF1 mRNA in breast cancer tissue,paracancerous tissue,and normal breast tissue;Western blotting method was used to detect the expression levels of TM4SF1 in breast cancer tissue,paracancerous tissue,and normal breast tissue;RT-PCR method was used to detect the expression levels of TM4SF1 mRNA in breast cancer tissue,paracancerous tissue, and normal breast tissue.The positive expression rates of TM4SF1 in breast cancer tissue of the breast cancer patients with different clinicopathological features were detected.Results:The positive expression rate of TM4SF1 in the breast cancer tissue was significantly higher than those in paracancerous tissue and normal breast tissue (P <0.05);there was no significant difference in the positive expression rates of TM4SF1 between paracancerous tissue and normal breast tissue (P = 0.531);the expression of TM4SF1 was not correlated with age,but was closely correlated with tumor size,differentiation degree,lymph node metastasis and tumor stage (P <0.05);the positive expression rate of TM4SF1 in basal like breast cancer tissue was higher than those in the other three types of tissues (P <0.05).The results of Western blotting showed that the expression level of TM4SF1 in breast cancer tissue was higher than those in paracancerous tissue and normal breast tissue (P < 0.05 ), but there was no significant difference in the expression level of TM4SF1 between paracancerous tissue and normal breast tissue (P >0.05). The results of RT-PCR showed that the expression level of TM4SF1 mRNA in breast cancer tissue was higher than those in the paracancerous tissue and normal breast tissue (P <0.01);there was no significant difference in the expression level of TM4SF1 mRNA between paracancerous tissue and normal breast tissue (P > 0.05 ). Conclusion:The TM4SF1 is highly expressed in breast cancer tissue. TM4SF1 may affect the occurrence, development and distant metastasis of breast cancer through various mechanisms.TM4SF1 may be a potential target for the treatment of breast cancer.