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Objective To investigate the expression level of Annexin Ⅱ in bronchial lung cancer tissue,alveolar lavage fluid(BALF) and pleural effusion and its value in the diagnosis of lung cancer.Methods From January 2014 to March 2016 in Jingzhou Infectious Disease Hospital,80 cases of patients with pleural effusion of lung cancer (Lung Cancer Group),40 patients with benign pleural effusion (benign group) were chose.The expression level of Annexin of two groups of patients with pleural effusion,BALF were detected and compared.Sixty specimens of department of pathology past (after lung cancer surgery cancer) and 30 cases of previously collected normal lung tissues (normal lung tissue) were collected,differential expression by immunohistochemical staining were compared between the two groups of Annexin Ⅱ protein.Results The positive expression of Annexin Ⅱ in lung cancer was 61.67％ (37/60),higher than that in normal lung tissue of 23.33％ (7/30),the difference was statistically significant(P＜0.001).The positive expression of Annexin Ⅱ in lung cancer patients was significantly correlated with rate of lymph node metastasis and clinical stage of lung cancer(P =0.039,0.029).Group of patients with pleural effusion,BALF expression of Annexin Ⅱ the level was higher than that in benign group(P＜0.001).When the critical value of Annexin in pleural effusion of lung cancer diagnosis was 10.94 μg/L,the sensitivity of diagnosis of lung cancer was 83.47％,the specificity was 80.66％,AUC value was 0.837.The critical value when BALF Annexin Ⅱ in the diagnosis of lung cancer was 52.08 μg/L,the sensitivity of diagnosis of lung cancer was 86.19％,the specificity was 89.22％,the AUC value was 0.898.Conclusion The expression level of Annexin Ⅱ in the BALF and pleural effusion of patients with bronchial lung cancer and lung cancer is increased,and it has a certain clinical value for the differential diagnosis of lung cancer.
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Objective To investigate the expression levels of Ezrin and AnnexinⅡ in gallbladder carci-noma and their association with clinicopathologic parameters and metastasis potential. Methods The tissue mi-croarray consisted of 59 gallbladder carcinoma tissues and 6 normal gallbladder tissues were examined for the ex-pression of Ezrin and AnnexinⅡusing immunohistochemistry technique. The expression of Ezrin and AnnexinⅡin 20 cases of fresh gallbladder carcinoma and 6 cases of normal gallbladder were measured with western blot. Results The expression of Ezrin and AnnexinⅡ were higher in the gallbladder cancer than those in the normal gallbladder tissue. The positive rate of Ezrin and AnnexinⅡ were 47.5% and 50.8% respectively. The expression of Ezrin was significantly correlated with live metastasis , lymph node metastasis and Nevin stages. The expression of AnnexinⅡwas significantly correlated with live metastasis , differentiation levels and Nevin stages. The expres-sion of Ezrin was correlated with AnnexinⅡ. Results of western blot suggested that Ezrin and Annexin II were highly expressed in gallbladder carcinoma tissues. The high expression of Ezrin and Annexin is closely related with liver invasion. Conclusion Measurement of the expression of Annexin and Ezrin II have important clinical significances to evaluate the malignant biological behavior of gallbladder carcinoma.
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Objective To establish a SYBR Green based real-time fluorescence quantitative PCR method for detecting human Annexin Ⅱ mRNA expression,and to detect the level of Annexin Ⅱ mRNA in human breast cancer cells MCF-7 and MDA-MB-231.Methods The specific primers were designed according to the conserved sequence of human Annexin Ⅱ gene.Total RNAs were extracted from human breast cancer cells(MCF-7,MDA-MB-231),then RNAs were transcribed reversely into cDNAs.The plasmid standards were constructed.The relative expression levels of human Annexin Ⅱ mRNA in human breast cancer cells were detected by this method.Results The square(r2 )of correlation coefficient of the standard curve in this method was 0.997,the melting curve analysis showed the single peak.The the intra-batch and inter-batch variable coefficients in the pGM-T Annexin Ⅱplasmid standard substance were 6.2%,7.8% and 9.1%,12.3% respectively.The further study indicated that AnnexinⅡ mRNA in MDA-MB-231 was higher than that in MCF-7(P<0.01).Conclusion The established SYBR Green real time fluorescence quan-titative PCR method for detecting human AnnexinⅡ is of good specificity and repeatability and can be used for quantitatively detec-ting AnnexinⅡ mRNA in breast cancer cells.
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Objective To explore the role of Annexin Ⅱ and urokinase-type plasminogen activator(uPA) proteins in the development and lymph node metastasis of thyroid cancer.Methods Immunohistochemistry was used to detect the expression of Annexin Ⅱ and uPA proteins in 35 cases of papillary thyroid cancer tissues,10 cases of thyroid adenoma tissues,16 cases of nodular goiter tissues and 7 cases of normal thyroid tissues.Results The positive expression rate of Annexin Ⅱ and uPA protein was significantly higher in papillary thyroid cancer than in normal thyroid tissues,nodular goiter tissues and thyroid adenoma tissues( P ＜0.05 ).The expression of Annexin Ⅱ and uPA protein was positively correlated in papillary thyroid cancer tissues(P ＜0.05).The expression of Annexin Ⅱ protein in thyroid cancer tissues was associated with tumor size and lymph node metastasis while the expression of uPA protein was only related to lymph node metastasis.Conclusions Annexin Ⅱ and uPA proteins are involved in the occurrence and development of papillary thyroid cancer.They also play a facilitating role in lymph node metastasis of papillary thyroid cancer.
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Objective To explore the expression and significance of Annexin Ⅱ protein in ovarian adenocar-einoma.Methods The expressions of Annexin Ⅱ protein were detected in 48 cases of ovarian adenocarcinoma and 10 cases of ovarian adenoma by SP immunohistochemistry.Results The positive rates of expression of Annexin Ⅱprotein in ovarian adenoma were 50.00% (5/10), and were 81.25% (39/48) in ovarian adenocareinoma (χ2 =4.414, P <0.05), in well and moderately-differentiated, poorly differentiated ovarian serous adenocarcinoma were 93.33% (14/15) and 78.95% (15/19) respectively (χ2 = 1.383 ,P0.05 ), and on the Ⅰ and Ⅱ stage, Ⅲ and Ⅳ stage of ovarian serous adenocarcinoma were 60.00% (6/10) and 95.83% ( 23/24 ) respectively (χ2=7.226, P <0.05).The positive rate in the group of ovarian serous adenocarcinoma with lymph node metastasis was 90.00% (27/30), while 50.00% (2/4) in the group without lymph node metastasis (χ2=4.502, P<0.05).Conclusion The positive expression of Annexin Ⅱ protein may be correlated with the carcinogenesis, development and lymph node metastasis of ovarian serous adenocarcinoma.
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Objectives:To investigate the effects of small interference RNA(siRNA) silencing Annexin Ⅱ on invasion and metastasis of human breast cancer cell MDA-MB-231.Methods:The chemically-synthetic siRNA duplexes targeting Annexin Ⅱ(Annexin Ⅱ-siRNA) was transiently transfected into MDA-MB-231 cells which have high metastatic potential by lipofectamin 2000.The transfection efficiency was observed by fluorescence microscopy.RT-PCR and western blot were used to semi-quantify the Annexin Ⅱ mRNA and protein levels.The malignant characters of transfected MDA-MB-231 cells including celluar proliferation rate,the activities of invasion and metastasis were analyzed by use of 3-(4,5-dimethylthiazol-2-yl)-5-diphenyl tetrazolium bromide(MTT) and millicell chamber assay,respectivilly.Results:Annexin Ⅱ-siRNA effectively inhibited Annexin Ⅱ mRNA and protein levels(P
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Objective To explore the effects of antisense vector of annexinⅡ gene on the growth of SPC A 1, a human lung cancer cell line. Methods The total RNA was isolated from human lung cancer cell line SPC A 1 and the target DNA fragments were amplified by RT PCR. The antisense expression vector was constructed by double restriction endonuclease cleavage directional clone method. Annexin Ⅱ antisense expression vector was introduced into SPC A 1 cells by liposome transfection reagent. The expression of annexin Ⅱ mRNA was analyzed by semi quantitative RT PCR. The effects of antisense vector of annexinⅡ gene on the growth of SPC A 1 were observed. Results The antisense vector of annexinⅡ gene was constructed and introduced into SPC A 1 cells successfully. Semi quantitative RT PCR showed that the annexin Ⅱ mRNA expression reduced by about two thirds in the transfected cells as compared with that in the untransfected cells. Compared with the untransfected cells, transfected cells decreased significantly in cell growth, clone formation efficiency in plating and DNA synthesis. Cell cycle was blocked in G 0 G 1 phase. Conclusion Annexin Ⅱ could promote the growth of lung cancer cells and may be helpful for the development of lung cancer.
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Objective To explore the annexinⅡexpression in the differentiation of human epidermal keratinocytes. Methods Immunohistochemistry was used to detect annexinⅡexpression in early, middle, late human embryonic skin and adult skin, respectively. Results The expression of annexinⅡ in embryonic epidermis was significantly increased with the development of embryo and annexin Ⅱ expressed much higher in adult epidermis than in embryonic epidermis. Annexin Ⅱ localized in the membrane of keratinocyte, mainly in suprabasal, the more differentiated layers of the epidermis. Conclusion Annexin Ⅱ may be involved in regulating the differentiation of human epidermal keratinocytes.
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Objective In order to make further investigation of the functional meaning of Ca~(2+)dependent phospholipids binding protein AnnexinⅡ,we tested the effects of AnnexinⅡ combined with embryonic neural stem cells(NSCs) transplantion on the repair of spinal cord injury(SCI). Methods The spinal cord of the adult rats was transected completely between T_9-T_(10).AnnexinⅡ and NSCs were injected at the transected site.The lesion area of the spinal cord,growth of axon,and survival number and migration of the transplanted NSCs were measured.The survival number was shown by prelabeling the NSCs with Hoechest 33342 and the growth of axon traversing the transected area was shown by fluorogold(FG) retrograde tracing.The AnnexinⅡ injection+NSCs implantation groups were compared with groups that respectively received 1.NSCs implantation alone;2.sham-operation+NSCs implantation and 3.Vehicle injection of culture medium. Results Our results demonstrated that AnnexinⅡ treatment in vivo could significantly reduce the lesion area(P
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Objective To analyze the effects of annexin Ⅱ antisense expression vector on the growth of the transplanted tumor in nude mice. Methods The SPC-A-1 cells (parental group) and SPC-A-1-annexin Ⅱ cells (antisense group) were inoculated subcutaneously in nude mice respectively. The forming time, volume, and weight of tumor were measured. The tumor cells and the surrounding tissues were observed microscopically. Results The forming time of tumor in the antisense group was later than that in the parental group. The volume and weight of tumor in the antisense group were lower than those in the parental group. Furthermore, invasion of tumor in the surrounding tissues was found in 2 cases in the parental group, but not in the antisense group. Conclusion The annexin Ⅱ gene may be important in promoting tumor cell growth and invading the surrounding tissues.