The role of SALL4 in tumorigenesis, development and its significance in diagnosis and therapy / 中国肿瘤生物治疗杂志

@#[摘 要] 人类婆罗双树样基因4（spalt-like transcription factor 4，SALL4）是一种新发现的癌胚基因，也是维持胚胎干细胞自我更新和多向分化潜能的锌指蛋白转录因子。SALL4基因的表达随发育成熟的过程逐渐下调甚至在大多数正常的成体组织中不表达。然而，近年来的研究表明，SALL4在多种肿瘤包括实体肿瘤和血液系统肿瘤中异常高表达，且SALL4表达水平与肿瘤发生发展及肿瘤患者的不良预后、生存期相关，因而成为肿瘤预测的生物学标志。由于SALL4表达模式的特异性及其在肿瘤发生发展中的作用使其成为肿瘤治疗的潜在靶点。本文就SALL4在肿瘤发生发展中的作用、调控肿瘤发生发展的机制及SALL4在疾病诊疗中的意义等方面作一综述。

Effect of SALL4 on radiosensitivity of leukemia cell line HL-60 / 中华放射肿瘤学杂志

Objective To evaluate the effect of down-regulating SALL4 on the radiosensitivity of leukemia cells,aiming to provide new ideas for improving radiosensitivity of leukemia patients.Methods Human acute myeloid leukemia cell line HL-60 infected with shRNA SALL4 and shRNA control lentivirus was classified into the Lv-shSALL4 group and Lv-shNC group.The levels of SALL4 mRNA and protein in cells were detected by RT-PCR and Western blot.The infected cells treated with 8 Gy dose irradiation were assigned into the Lv-shSALL4 + radiation and Lv-shNC+radiation groups.The cell apoptosis was detected by flow cytometry.The levels of cleaved Caspase-3,cleaved Caspase-9 and Bax proteins in cells were determined by Western blot.The cells in the Lv-shSALL4 and Lv-shNC groups were exposed to 0,2,4,6 and 8 Gy irradiation.The radiosensitivity ratio was determined by cell clone test.Results The level of SALL4 in the Lv-shSALL4 group was significantly lower than that in the Lv-shNC group (P＜0.05).The cell apoptosis rate was significantly increased,the levels of cleaved Caspase-3,cleaved Caspase-9 and Bax proteins were remarkably up-regulated in cells compared with those in the Lv-shNC group (all P＜ 0.05).The cell proliferation ability in the Lv-shSALL4 + radiation was significantly reduced,the cell apoptosis rate was considerably increased,the levels of cleaved Caspase-3,cleaved Caspase-9 and Bax proteins were significantly up-regulated compared with those in the Lv-shSALL4 and Lv-shNC+ radiation groups (all P＜0.05).The cell radiosensitization ratio in the Lv-shSALL4 group was 1.323.Conclusion Down-regulating SALL4 can increase the radiosensitivity of leukemic cells,inhibit the cell proliferation and induce the apoptosis of leukemic cells.

Value of Sal-like 4 in the diagnosis and treatment of primary liver cancer / 临床肝胆病杂志

Sal-like 4 (SALL4) is a transcription factor that helps to maintain self-renewal and pluripotency of stem cells. This gene is expressed in the human fetal liver, but is silent in the healthy adult liver. It has been reported that SALL4 has a diagnostic value in a variety of solid tumors. These characteristics make SALL4 a possible tumor biomarker for liver cancer. This article reviews the recent advances in the role of SALL4 in the development, progression, metastasis, and other biological processes of primary liver cancer, as well as its role in predicting the prognosis of primary liver cancer. We believe that with further studies, SALL4 and related microRNAs may become new markers for the diagnosis, treatment, and prognostic evaluation of primary liver cancer.

Effect of SALL4 on radiosensitivity of leukemia cell line HL-60 / 中华放射肿瘤学杂志

Objective@#To evaluate the effect of down-regulating SALL4 on the radiosensitivity of leukemia cells, aiming to provide new ideas for improving radiosensitivity of leukemia patients.@*Methods@#Human acute myeloid leukemia cell line HL-60 infected with shRNA SALL4 and shRNA control lentivirus was classified into the Lv-shSALL4 group and Lv-shNC group. The levels of SALL4 mRNA and protein in cells were detected by RT-PCR and Western blot. The infected cells treated with 8 Gy dose irradiation were assigned into the Lv-shSALL4+ radiation and Lv-shNC+ radiation groups. The cell apoptosis was detected by flow cytometry. The levels of cleaved Caspase-3, cleaved Caspase-9 and Bax proteins in cells were determined by Western blot. The cells in the Lv-shSALL4 and Lv-shNC groups were exposed to 0, 2, 4, 6 and 8 Gy irradiation. The radiosensitivity ratio was determined by cell clone test.@*Results@#The level of SALL4 in the Lv-shSALL4 group was significantly lower than that in the Lv-shNC group (P<0.05). The cell apoptosis rate was significantly increased, the levels of cleaved Caspase-3, cleaved Caspase-9 and Bax proteins were remarkably up-regulated in cells compared with those in the Lv-shNC group (all P<0.05). The cell proliferation ability in the Lv-shSALL4+ radiation was significantly reduced, the cell apoptosis rate was considerably increased, the levels of cleaved Caspase-3, cleaved Caspase-9 and Bax proteins were significantly up-regulated compared with those in the Lv-shSALL4 and Lv-shNC+ radiation groups (all P<0.05). The cell radiosensitization ratio in the Lv-shSALL4 group was 1.323.@*Conclusion@#Down-regulating SALL4 can increase the radiosensitivity of leukemic cells, inhibit the cell proliferation and induce the apoptosis of leukemic cells.

Clinicopathological characteristics of gastric cancer with enteroblastic differentiation / 中国肿瘤临床

Objective: To investigate the clinicopathological features and molecular phenotypes of gastric cancer with enteroblastic dif-ferentiation (GCED). Methods: A retrospective analysis of 337 patients with gastric adenocarcinoma diagnosed by the pathology de-partment of the First Affiliated Hospital of Zhejiang University in March 2013-2017 was conducted. Of them, 8 patients were diag-nosed with gastric carcinoma with intestinal blastocyte differentiation. All the patients were elderly, including 6 men and 2 women. The onset age was 68-83 years (mean 76.6 years). Two cases had serum AFP≥200 μg/L before treatment. According to the histopatho-logical morphology, the immunophenotype was analyzed by immunohistochemistry, the SALL4 gene was detected using reverse tran-scription-polymerase chain reaction (RT-PCR), and the relevant literature was reviewed. Results: Microscopically, all cases had primi-tive enteroid structures, consisting of cubic or columnar cells with clear cytoplasm, and immunohistochemical staining showed positivi-ty for either AFP and GPC3 or SALL4. The expression of SALL4 mRNA was significantly increased by RT-PCR. Follow-up from 1 to 5 years showed that 5 patients had liver and other organ metastases, 2 patients died, and 1 patient survived without a tumor. Conclusions:GCED is a rare invasive gastric adenocarcinoma with a worse prognosis than that of normal intestinal adenocarcinoma. The treatment of general intestinal adenocarcinoma has little effect. There are some characteristic changes in histology. It would be helpful for diag-nosis and differential diagnosis if clinicians are familiar with the tumor spectrum and genetic characteristics. Target therapy for an origi-nal marker, such as SALL4, has a bright future.

Positive immunostaining of Sal-like protein 4 is associated with poor patient survival outcome in the large and undifferentiated Korean hepatocellular carcinoma

PURPOSE: Previous studies have shown the role of Sal-like protein 4 (SALL4) as a biomarker in hepatocellular carcinoma (HCC), and some studies have shown the relationship between SALL4 and prognosis. Given the debates in study groups differences in terms of etiologic causes between Western and Asian HCC and detection methods, we attempted to verify the features of SALL4 immunoreactivity and its clinical correlation in Korean HCC patients. METHODS: Immunohistochemical staining of SALL4 of tissue microarrays (TMAs) consisting of 213 surgically resected HCC patients' tissue were scored in a semiquantitative scoring system with immunoreactive score and the results analyzed with clinical outcome, in addition to general demographics and clinical characteristics. RESULTS: SALL4 immunoreactivity was expressed in 50 cases. Relevance between SALL4 and α-FP correlated significantly (P = 0.002). Also, the SALL4-positive patients had considerably higher tumor grade (P < 0.001). The survival analysis showed negative correlation with SALL4 immunoreactivity in all HCC patient groups, but SALL4 immunoreactivity in T3 and T4 HCC correlated with poor prognosis. CONCLUSION: Here, we found that positive immunostaining of SALL4 is correlated with poor patient survival outcome in large and undifferentiated Korean HCC. SALL4 expression showed close relationship with clinical outcomes of HCCs in Korean patients.

Positive immunostaining of Sal-like protein 4 is associated with poor patient survival outcome in the large and undifferentiated Korean hepatocellular carcinoma

PURPOSE: Previous studies have shown the role of Sal-like protein 4 (SALL4) as a biomarker in hepatocellular carcinoma (HCC), and some studies have shown the relationship between SALL4 and prognosis. Given the debates in study groups differences in terms of etiologic causes between Western and Asian HCC and detection methods, we attempted to verify the features of SALL4 immunoreactivity and its clinical correlation in Korean HCC patients. METHODS: Immunohistochemical staining of SALL4 of tissue microarrays (TMAs) consisting of 213 surgically resected HCC patients' tissue were scored in a semiquantitative scoring system with immunoreactive score and the results analyzed with clinical outcome, in addition to general demographics and clinical characteristics. RESULTS: SALL4 immunoreactivity was expressed in 50 cases. Relevance between SALL4 and α-FP correlated significantly (P = 0.002). Also, the SALL4-positive patients had considerably higher tumor grade (P < 0.001). The survival analysis showed negative correlation with SALL4 immunoreactivity in all HCC patient groups, but SALL4 immunoreactivity in T3 and T4 HCC correlated with poor prognosis. CONCLUSION: Here, we found that positive immunostaining of SALL4 is correlated with poor patient survival outcome in large and undifferentiated Korean HCC. SALL4 expression showed close relationship with clinical outcomes of HCCs in Korean patients.

Research progress of SALL4 gene in malignant tumor disease / 实用肿瘤学杂志

Malignant tumor is a serious threat to human health and its pathogenesis is complex , which caused by interaction of a variety of carcinogenic factors in the human body .SALL4 gene is a kind of newly dis-covered gene and it plays an important role in function of embryonic stem cell due to its protein transcription fac -tor with C2H2 zinc finger domain.Studies found that this gene mutation often results in occurrence of malignant tumor.This paper will summarize the molecular mechanism of SALL 4 in the occurrence and development of malig-nant tumor and its diagnosis and treatment on malignant tumor .

SALL4 Expression in Hepatocellular Carcinomas Is Associated with EpCAM-Positivity and a Poor Prognosis

BACKGROUND: There is increasing interest in hepatocellular carcinomas (HCC) expressing "stemness"-related markers, as they have been associated with aggressive behavior and poor prognosis. In this study, we investigated the usefulness of Sal-like protein 4 (SALL4), a recently proposed candidate marker of "stemness." METHODS: Immunohistochemical stains were performed for SALL4, K19, and epithelial cellular adhesion molecule (EpCAM) on tissue microarrays constructed from 190 surgically resected HCCs, and the results were correlated with the clinicopathological features and patient survival data. RESULTS: Nuclear SALL4 expression was observed in 39/190 HCCs (20.5%), while K19 and EpCAM were expressed in 30 (15.9%) and 92 (48.7%) HCCs, respectively. The nuclear expression was generally weak, punctate or clumped. SALL4 expression was significantly associated with a poor overall survival compared to SALL4-negative HCCs (p = .014) compared to SALL4-negative HCCs. On multivariate analysis adjusted for tumor size, multiplicity, vascular invasion, and pathological tumor stage, SALL4 remained as a significant independent predictor of decreased overall survival (p= .004). SALL4 expression was positively correlated with EpCAM expression (p = .013) but not with K19 expression. HCCs that expressed both SALL4 and EpCAM were associated with significantly decreased overall survival, compared to those cases which were negative for both of these markers (p = .031). CONCLUSIONS: Although SALL4 expression was not significantly correlated with other clinicopathological parameters suggestive of tumor aggressiveness, SALL4 expression was an independent predictor of poor overall survival in human HCCs, and was also positively correlated with EpCAM expression.

Diagnostic utility of immunohistochemical markers SALL4, D2-40 and Glypican-3 in primary testicular germ cell tumors / 临床与实验病理学杂志

Purpose To investigate the diagnostic utility of the immunohistochemical markers SALL4, D2-40 and Glypican-3 in prima-ry testicular germ cell tumors (TGCTs). Methods The expression of SALL4, D2-40 and Glypican-3 protein was detected by EnVi-sion immunohistochemical method in 56 cases of primary testicular germ cell tumors, including 5 intratubular germ cell neoplasms ( IT-GCNs) , 10 seminomas, 14 embryonal carcinomas ( ECs) , 14 yolk sac tumors ( YSTs) , 1 choriocarcinoma, 5 immature teratomas and 12 mature teratomas. 10 normal testicular tissues and 5 lymphomas were selected as control. Results All of ITGCNs, seminomas, YSTs and ECs were diffusely strongly positive for SALL4. Focal SALL4 staining was seen in choriocarcinoma, 3 of 5 immature terato-mas and 3 of 12 mature teratomas. All of ITGCNs, seminomas showed diffusely strong D2-40 staining. ECs (4/14) were focally posi-tive for D2-40, while choriocarcinoma, YSTs and teratomas were negative for D2-40. Glypican-3 was diffusely positive in YSTs (13/14), and focally weakly positive in ECs (2/14), respectively. ITGCNs, seminomas, choriocarcinoma and teratoma were negative for Glypican-3. In contrast, 10 normal testicular tissues and 5 lymphomas showed no SALL4, D2-40 and Glypican-3 staining. Conclu-sions SALL4 is a useful diagnostic marker with high sensitivity and specificity for TGCTs. Combination of SALL4, D2-40 and Glypi-can-3 is helpful to the diagnosis and differential diagnosis for TGCTs.

Expression and clinical significance of SALL4 expression in gastric carcinoma tissues / 重庆医学

Objective To study the expression and clinical significance of the SALL4 in human gastric carcinoma tissues. Methods The expression of SALL4 in 91 samples of gastric carcinoma and 37 samples of normal gastric tissues was detected by RT-PCR,Western blot and immunohistochemistry,and its relationship with the clinical data were analyzed statistically.Results The positive expression rate of SALL4 in gastric carcinoma(74.7%)was significantly higher than that(18.9%)in normal gastric mucosa tissues(P <0.05).Moreover,with the decreased with the differentiation of gastric carcinoma,the positive expression rate of SALL4 was increased.The expression of SALL4 mRNA and protein in gastric carcinoma tissues were significantly higher than that in normal gastric tissues(P <0.050).The expression levels of SALL4 were relevant to lymph node metastasis(P =0.001),infiltra-tion depth(P =0.029)and the differentiation degree of gastric carcinoma(P =0.050).Conclusion SALL4 was highly expressed in gastric cancer tissues and relevant to lymph node metastasis,infiltration depth and the differentiation degree,which may have play an important role in the development of gastric cancer.

Knockdown of SALL4 expression regulates cell proliferation and apoptosis in prostate cancer LNCaP cells / 中国病理生理杂志

[ ABSTRACT] AIM: To investigate the SALL4 expression, proliferation and apoptosis in the LNCaP cells after transfection of SALL4 siRNA.METHODS: The expression of SALL4 at mRNA and protein levels was detected by real-time PCR and Western blotting.MTS assay, colony formation assay and flow cytometry were used to determine the prolifer-ation, colony formation ability and apoptosis of the LNCaP cells.The effect of SALL4 on the expression of Bax and Bcl-2 was analyzed by Western blotting.RESULTS:Compared with negative control group, the expression of SALL4 at mRNA and protein levels in LNCaP cells was down-regulated by transfection of SALL4 siRNA ( P<0.05 ) .The proliferation rate and colony formation ability were decreased, while apoptosis rate increased in si-SALL4 group (P<0.05).Higher expres-sion of Bax and lower expression of Bcl-2 in si-SALL4 group were observed ( P<0.05 ) .CONCLUSION:Down-regula-tion of SALL4 by siRNA not only suppresses LNCaP cell proliferation and colony formation, but also inhibits Bcl-2 expres-sion and activates Bax expression to induce apoptosis.

Study on expression of stem cell marker SALL4 gene in cervical cancer / 重庆医学

Objective To research the expression and clinical significance of SALL 4 gene in cervical cancer .Methods The ex-pression of SALL4 in 56 samples of cervical cancer and 35 samples of normal cervical tissues was detected by immunohistochemistry and RT-PCR ,and its relationship with the clinicopathological characteristics of cervical cancer was analyzed .Results The expres-sion of SALL4 mRNA was 2 .56 ± 0 .22 in cervical cancer tissues ,which was significantly higher than 0 .38 ± 0 .03 in the normal cer-vical tissues .the difference between them had statistical significance(t=58 .1 ,P0 .05) .Conclusion SALL4 is highly expressed in the cervical cancer tissues and correlated with the tumor differentiation ,which might play an important role in the occurrence and development of cervical cancer .

SALL4 expression in children with acute leukemia and its clinical significance / 中国肿瘤临床

Objective:To investigate the expression of sal-like 4 (SALL4) gene in children with acute leukemia and analyze its clinical significance. Methods:Real-time PCR and immunohistochemistry were used to detect SALL4 mRNA and SALL4 protein ex-pressions in 50 patients initially diagnosed with acute leukemia and in 15 patients with immune thrombocytopenic purpura (ITP), which served as controls. Changes were detected in SALL4 mRNA expression from preliminary diagnosis and after complete remission of 5 acute leukemia patients. The relationship between SALL4 mRNA expression and clinical indicators was analyzed. Results: SALL4 mRNA expression is higher in initially diagnosed B-ALL [13.89 (1.00-63.15)] and AML [11.12 (2.31-56.59)] than in ITP controls [1.00 (0.29-1.71)] (P0.05). SALL4 protein expression is in agreement with SALL4 mRNA expression. SALL4 mRNA expression significant-ly decreased in complete remission stage [0.98 (0.22-1.09)] than in acute phase [28.64 (11.20-87.46)] in acute-leukemia patients (P0.05). Conclusion:SALL4 was found to play an important role in pro-moting childhood B-ALL and AML, which promises a new target for monitoring the therapeutic effects and evaluating the prognosis of childhood B-ALL and AML.

Expression of SALL4, Bmi-l and β-catenin in esophageal squamous cell carcinoma and the related clinical significance / 第二军医大学学报

Objective To investigate the expression of SALL4, Bmi-1 and β-catenin in esophageal squamous cell carcinoma (ESCC) and the related clinical implications. Methods Immunohistochemical staining was used to detect the expression of SALL4, Bmi-1 and β-catenin in 70 normal esophageal mucosa specimens, 70 dysplasia mucosa specimens and 123 ESCC specimens; and the relationship of their expression with the clinicopathological characteristics of ESCC was analyzed. Results The positive rates of SALL4, Bmi-1 and the aberrant rate of β-catenin expression gradually increased in order in normal esophageal mucosa, dysplasia mucosa andESCC groups. The positive rates of SALL4, Bmi-1 and the aberrant rate of β-catenin expression in the dysplasia mucosa and ESCC groups were significantly higher than those in normal esophagealmucosa group (P0. 05). In the dysplasia mucosa group, the positive rate of Bmi-1 increased along with the degree of dysplasia (P<0. 01). In the ESCC cases, the positive rate of Bmi-1 and aberrant rate of β-catenin were corelated with depth of invasion, degree of differentiation and lymph node metastasis of ESCC (P<0. 05), and positive rate of SALL4 was correlated with the clinical staging (P<0. 05) and lymph node metastasis of ESCC (P<0. 01). The expression of SALL4, Bmi-1 and β-catenin in the 123 cases of ESCC were positively correlated with each other (SALL4 and Bmi-1: r = 0. 373, P<0. 01; SALL4 and p-catenin: r=0. 214, P<0. 05; Bmi-1 and p- catenin: r=0. 204, P<0. 05). Conclusion SALL4, Bmi-1 and β-catenin might be involved in the development, progression, invasion and metastasis of ESCC; and the three of them might interact through corresponding signal pathways.

Very small embryonic-like stem-cell optimization of isolation protocols: an update of molecular signatures and a review of current in vivo applications

As the theory of stem cell plasticity was first proposed, we have explored an alternative hypothesis for this phenomenon: namely that adult bone marrow (BM) and umbilical cord blood (UCB) contain more developmentally primitive cells than hematopoietic stem cells (HSCs). In support of this notion, using multiparameter sorting we were able to isolate small Sca1+Lin-CD45- cells and CD133+Lin-CD45- cells from murine BM and human UCB, respectively, which were further enriched for the detection of various early developmental markers such as the SSEA antigen on the surface and the Oct4 and Nanog transcription factors in the nucleus. Similar populations of cells have been found in various organs by our team and others, including the heart, brain and gonads. Owing to their primitive cellular features, such as the high nuclear/cytoplasm ratio and the presence of euchromatin, they are called very small embryonic-like stem cells (VSELs). In the appropriate in vivo models, VSELs differentiate into long-term repopulating HSCs, mesenchymal stem cells (MSCs), lung epithelial cells, cardiomyocytes and gametes. In this review, we discuss the most recent data from our laboratory and other groups regarding the optimal isolation procedures and describe the updated molecular characteristics of VSELs.

Induction of acute myelogenous leukemia cell apoptosis by bortezomib and its effect on SALL4 gene expression / 中国药学杂志

OBJECTIVE: To investigate the proteasome inhibitor bortezomib induces acute myelogenous leukemia cell lines TF1 and NB4 apoptosis and its effect on SALL4 gene expression. METHODS: Cell proliferations were analyzed by MTT assay. Flow cytometry was used to analyze cell apoptosis rate. SALL4 protein was detected by immunocytochemistry. The expressions of SALL4 gene were detected by Real-time PCR. SALL4 proteins in two cell lines were detected by Western Blotting. RESULTS: MTT assay showed bortezomib inhibited the proliferations of two cell lines in a time-and-does manner. TF1 and NB4 cells' 48 h IC50 were (29.15 ± 0.55) and (30.55 ± 0.74) nmol · L-1 respectively. Flow cytometry showed bortezomib could induce apoptosis of two cell lines in a dose-dependent manner. Immunocytochemistry analysis revealed that both of two cell lines expressed SALL4 proteins which located in cell nucleus. Real-time PCR demonstrated that SALL4 genes were down-regulated after cells were treated by different concentrations(10, 30, 50 nmol · L-1) of bortezomib for 24 h, and bortezomib 50 nmol · L-1 groups' genes were down-regulated to 45.11% (TF1) and 69.77% (NB4) respectively comparing with the control groups (P < 0.05). Western blotting revealed that both of the cell lines expressed SALL4B proteins and which could be inhibited by bortezomib in a time-and-does manner. CONCLUSION: Bortezomib can significantly inhibit two cell lines proliferation and induce apoptosis, meanwhile down-regulate the expressions of SALL4 gene. Copyright 2012 by the Chinese Pharmaceutical Association.

SALL4 and hematologic neoplasms / 国际肿瘤学杂志

SALL4 gene is closely related to body malformations related diseases, embryonic stem cell development, and hematopoietic malignancies. SALL4 can activate hematopoietic stem cell through Wnt signaling pathways, and promote continued proliferation of leukemia stem cells, leading to leukemia. In-depth study of SALL4 gene and its protein function will help clarify the pathogenesis of leukemia, providing a new target for the treatment of leukemia.

Construction of SALL4 shRNA vectors and THP-1 cell transfection / 白血病·淋巴瘤

Objective To construct an efficient SALL4 shRNA vector and transfect it into THP-1 cells for investigating the effect of SALL4 in leukemia. Methods Four SALL4-specific siRNA to aim at different SALL4 mRNA target sites and a negative control siRNA were designed, and pGPU6/GFP/Neo/SALL4 shRNA vectors were constructed. THP-1 cells were transfected and the expression of SALL4 in shRNA detected, and blank control and negative control were also designed. Results The results of real time quantitive PCR and Western blotting both exhibited that the interference effect of pGPU6/GFP/Neo/SALL4 shRNA-B vector was optimal targeting to mRNA-1122 target site and down-regulated the expression of SALL4 more significant(P <0.05). Conclusion Successfully construction of SALL4 siRNA vector by choosing SALL4 shRNA-B would be useful to accomplish study of SALL4.