Caracterización de marcadores de células madre tumorales en cáncer escamocelular de cavidad oral

Introducción: El cáncer escamocelular de cavidad oral es una patología con bajas tasas de sobrevivencia. Cuando no es tratado adecuadamente es un tumor de alta recurrencia y resistente al tratamiento. Nuevas hipótesis plantean que las células tumorales progenitoras por sus propiedades de auto renovación, iniciación tumoral, migración y metástasis pueden ser responsables de la manutención y renovación de este tumor. Sin embargo, aún no existe un consenso sobre la verdadera participación de ellas, debido a que su identificación y caracterización es aún un reto experimental. Objetivo: En este trabajo se busca detectar células con expresión de marcadores de células tumorales Progenitoras en muestras cáncer escamocelular de cavidad oral y relacionarlo con los estadios de diferenciación del tumor. Metodología: En esta investigación se tomaron 32 muestras de pacientes con carcinoma escamocelular de cavidad oral. Se logró detectar in situ, mediante la técnica de inmunofluorescencia, cuatro reconocidos marcadores de células tumorales progenitoras. Resultados: Se identificaron los marcadores OCT4, SSEA4, NANOG y TRA-1-60 en los diferentes estadios de diferenciación tumoral, lo que sugiere la participación de las células progenitoras tumorales en la evolución de esta patología. Conclusiones: El establecimiento y correcta identificación de las células tumorales progenitoras abre nuevas vías terapéuticas para el abordaje de este tumor, en busca de mejorar el pronóstico, tasa de sobrevivencia y calidad de vida del paciente.

Introduction: Squamous cell carcinoma of the oral cavity is a pathology with poor survival rates. When it is not adequately treated, it is a tumor with high recurrence and resistance to treatment. According to new hypotheses, progenitor tumor cells, due to their properties of self-renewal, tumor initiation, migration, and metastasis, could be responsible for the maintenance and renewal of this tumor. However, there is still no consensus on their true participation, subsequent to difficult in their identification and characterization. Materials and methods: In this research, 32 samples provided from patients diagnosis with squamous cell carcinoma of the oral cavity were used. To detect specific markers progenitor tumor cells were used immunofluorescence microscopy. Results: The cells markers OCT4, SSEA4, NANOG and TRA-1-60 were identified in the different stages of the tumor samples, all these findings suggest the role of tumor progenitor cells in the evolution of this pathology. Conclusions: The establishment and correct identification of the progenitor tumor cells provide new therapeutic options for the approach of this tumor seeking to improve the prognosis, survival rate and quality of life of the patient.

The prognostic effect of immunohistochemical staining rates in patients with non–muscle-invasive bladder cancer

Context: Despite the follow-up protocols developed in non–muscle-invasive bladder cancer patients, progression and recurrence could not be prevented. Aims: We aimed to investigate whether proteins such as OCT-4, CD47, p53, Ki-67, and Survivin, which increase in bladder cancer cells, can be used as prognostic markers for patients with non–muscle-invasive bladder cancer. Settings and Design: The study included a total of 89 patients with newly diagnosed non–muscle-invasive bladder cancer between January 2015 and December 2020. Materials and Methods: Levels of OCT-4, CD47, p53, K?-67, and Survivin proteins in cancer cells were determined with a semi-quantitative immunohistochemical experiment. Pathological data and survival rates were compared according to the staining rates. Statistical Analysis Used: Data obtained in the study were analyzed statistically with SPSS 22.0 (SPSS, Chicago, IL, USA). Results: The mean age of the patients was 64.25 ± 9.91 years, and the median follow-up period was 55 months. Recurrence rate was determined to be 36% (n = 32), and the rate of progression at 40.4% (n = 36). The staining rates were stronger for each marker in the progression group and advanced-stage tumors (p < 0.001). The findings of the multivariate analysis carried out as part of the study showed that older age and higher tumor stage were independent risk factors for recurrence-free survival (HR = 1.048 and 7.074, respectively; P = 0.02). Also, higher tumor stages, diameters, and grades were associated with reduced progression-free survival (HR = 0.105, 0.395, 0.225, respectively; P < 0.05). Conclusions: Although immunohistochemical staining rates are promising, it is more appropriate to use tumor characteristics when assessing survival rate in patients with non–muscle-invasive bladder cancer.

Prognostic value of NOTCH1 and OCT4 in gastric carcinoma

Background: NOTCH1 pathway activation has been recently described to be a key player in gastric carcinogenesis, enhance the survival and proliferation of cancer stem cells (CSCs) and mediate chemoresistance in several malignancies. Aim: This study investigated the correlation between NOTCH1 and CSC marker OCT4 (octamer binding transcription factor-4) expression and the clinicopathological properties, survival and treatment outcome in patients with gastric carcinoma (GC) receiving adjuvant chemotherapy. Materials and Methods: NOTCH1 and OCT4 were immunohistochemically detected in 50 post-operated specimens of GC. Patients' data regarding disease-free survival (DFS), overall survival (OS), and the response to the chemotherapy was statistically analyzed. Results: NOTCH1 and OCT4 overexpression was detected in 60% and 52% of GC tissues, respectively, and that was significantly higher than the rates in adjacent non-neoplastic gastric mucosa (P < 0.05). A significant correlation was detected between overexpression of NOTCH1 and OCT4 in GC and aggressive clinicopathological features; poor differentiation (P = 0.021, P = 0.037, respectively), depth of tumor invasion (P < 0.001 for both), TNM stage (P < 0.001 for both), lymph node metastasis (P = 0.002, P = 0.003, respectively) and distant metastasis (P < 0.001 for both). NOTCH1 was positively correlated with OCT4 (P = 0.002). Survival analysis disclosed that upregulation of NOTCH1 and OCT4 was associated with worse DFS (P = 0.013, P < 0.001, respectively) and OS (P < 0.001 for both). Overexpression of NOTCH1 and OCT4 correlated with poor response to chemotherapy (P = 0.013, P = 0.005, respectively) and worse clinical outcome. Conclusion: Combined detection of these proteins might disclose even better predictive value for shorter survival and resistance to chemotherapy.

Study on the effects and mechanism of Linc01278 on the biological behavior of prostate cancer cells / 中华内分泌外科杂志

Objective:To investigate the effect and molecular mechanism of long non-coding RNA (Linc01278) on the proliferation, invasion and migration, and tumor phenotype of prostate cancer cells.Methods:Prostate cancer tissues and corresponding normal tissues adjacent to the cancer were obtained in 46 patients with prostate cancer confirmed by the hospital from Dec. 2018 to Dec. 2019. Real-time fluorescence quantitative PCR (qPCR) was used to detect the expression levels of Linc01278 and miR-145 in prostate cancer tissues and adjacent normal tissues. The prostate cancer PC-3 cells were transfected with plasmids and cells were divided into four groups: blank group, control group, overexpression group and rescue group. The blank group was not given any treatment; the control group was transfected with blank control vector and miRNA Control; the overexpression group was transfected with Linc01278 overexpression vector and miRNA Control; the rescue group was transfected with Linc01278 overexpression vector and miR-145 mimics. Bioinformatics analysis was used to predict the binding site of Linc01278 to miR-145. Dual Luciferase Reporter Gene Assay was used to analyze the adsorption of Linc01278 on miR-145; Transwell experiment was used to detect the invasion and migration ability of the four groups of cells; CCK-8 method was used to detect the cell proliferation ability of the four groups of cells. Western blot was used to detect the expression of OCT4 and SOX2 in the four groups of cells; qPCR was used to detect the expression of OCT4, SOX2 and miR-145 in the four groups of cells.Results:Compared with adjacent normal tissues, Linc01278 was significantly higher in prostate cancer tissues (adjacent normal tissues: 0.012±0.002 vs prostate cancer tissues: 0.022±0.002, P=0.0072) , while miR-145 was significantly lower (adjacent normal tissues: 0.117±0.011 vs prostate cancer tissues: 0.081±0.007, P=0.0005) . Correlation of both was negative significantly ( P=0.0012) . Targetscan analysis revealed that the 804-825 position of the Linc01278 transcript had a complementary base pairing with miR-145. The results of the double luciferase reporter gene showed that miR-145 mimics could significantly reduce the luciferase activity of Linc01278 ( P<0.01) . Compared with the blank group and the control group, the invasion and migration cells, the relative proliferation ability, the expression of Oct4 and Sox2 mRNA and protein were significantly increased, and the expression of miR-145 was significantly decreased in the overexpression group ( P<0.05) . While compared with the overexpression group, the invasion and migration cells, the relative proliferation ability, Oct4 and Sox2 were significantly decreased in the rescue group, and the expression of miR-145 was significantly increased ( P<0.01) . Conclusion:Linc01278 may promote the proliferation, invasion and migration of prostate cancer cells by specifically adsorbing miR-145.

Elevated expression of stemness genes in adipose-derived mesenchymal stem cells cultured on fibrin scaffold

In regenerative medicine, MSCs need to be pluripotent for better results. In this study, the effect of fibrinscaffold on expression of stemness genes was examined. Adipose-derived MSCs were cultured in tissue cultureplates (2D) and 3-dimensional (3D) fibrin scaffolds. The effect of fibrin scaffold on proliferation of adiposederived MSCs was evaluated by MTT assay. The expression of stemness genes (OCT4 and SOX2) wereevaluated by qRT-PCR, and flow cytometry was done for Nanog protein level. Cultured MSCs on fibrinscaffold were able to proliferate according to data obtained by MTT assay. Expression of OCT4 and SOX2 hada significant increase in cells were cultured in 3D condition compared to 2D condition (P \0.05). Also,increased expression of Nanog protein in 3D culture was observed (P \0.05). OCT4 and SOX2 in 3Dcondition increased two-fold and three-fold respectively in 2D and 3D conditions. Moreover, expression ofNanog increased 30% more than in 2D condition. Evaluation of important pluripotency regulators such asOCT4, SOX2, and Nanog showed that fibrin scaffolds are useful instruments to maintain stemness of MSCs,which is essential in field of stem cell therapy and regenerative medicine.

Suppression of OCT4-pg5 and OCT4B enhanced the sensitivity of bladder cancer cells T24 to cisplatin / 解放军医学杂志

Objective: To investigate the effects of octamer-binding transcription factor 4 pseudogene 5 (OCT4-pg5) and octamer-binding transcription factor 4B (OCT4B) expression on cisplatin sensitivity of bladder cancer T24 cells. Methods: The expression of OCT4-pg5 and OCT4B in bladder cancer cells and tissues were quantified by real-time quantitative reverse transcriptase-polymerase chain reaction (RT-qPCR). To examine the effect of OCT4-pg5 and OCT4B on cisplatin sensitivity, cells were transfected with OCT4-pg5 si-RNA, OCT4B si-RNA and their control si-RNA by lipofectamine 2000 followed by a 48 h cisplatin treatment. CCK8 was used to assess the half-maximal inhibitory concentration (IC50) of cisplatin in T24 cells. OCT4-pg5 and OCT4B expression were detected by RT-qPCR or Western blot. Next, flow cytometry was used to examine the effects of OCT4-pg5 and OCT4B on T24 cells apoptosis and cell-cycle distribution. Results: RT-qPCR results showed that OCT4-pg5 and OCT4B were highly expressed in bladder cancer cells and tissues. Moreover, OCT4-pg5 expression was positively correlated with OCT4B expression in 23 bladder cancer tissues. Cells transfected with OCT4-pg5 si-RNA or OCT4B si-RNA showed a much lower IC50 of cisplatin, compared with cells in the control group. RT-qPCR or Western blot results showed OCT4-pg5 or OCT4B expression was significantly down-regulated after being transfected with OCT4-pg5 si-RNA or OCT4B si-RNA in cisplatin treated T24 cells. The flow cytometry results showed that OCT4-pg5 suppression or OCT4B suppression of cisplatin treated T24 cells resulted in a significant increase of cell apoptosis and marked the transition from the S phase to G0/G1 phase. Conclusions Downregulating OCT4-pg5 and OCT4B could enhance the sensitivity of bladder cancer T24 cells to cisplatin.

Biological characteristics of human bone marrow mesenchymal stem cells in the microenvironment of rabbit bladder acellular matrix graft / 西安交通大学学报(医学版)

Objective: To explore the effects of the microenvironment of rabbit bladder acellular matrix graft (BAMG) on proliferation, cell surface markers, and molecular protein level of human bone marrow mesenchymal stem cells (hBMSCs). Methods: We prepared BAMG immersion fluid medium and detected its effect on the proliferation of hBMSCs by MTT method. The expressions of CD44, CD45, CD73 and PDGFRβ were detected by flow cytometry. The expressions of PPAR, OCN and α-SMA were detected by RT-PCR, and the expression of OCT was detected by Western blot. Results: hBMSCs had good compatibility with BAMG. The MTT method showed that BAMG and BAMG immersion medium did not affect the proliferation capacity of hBMSCs. The surface of hBMSCs cells cultured with immersion fluid still expressed CD44, CD73 and PDGFRβ, but not CD45. RT-PCR showed that OCN, PPAR, and α-SMA were all expressed. Western blot test also showed the positive expression of OCT-4. Conclusion: hBMSCs can still keep their original biological characteristics in the microenvironment of rabbit BAMG. It can be the seed cells and combined substrate materials for urinary system tissue engineering.

Isolation and identification of Oct4 RNA-binding proteins / 上海交通大学学报（医学版）

Objective: To identify the RNA binding proteins of Oct4, a key factor of embryonic stem cells. Methods: Total RNA was extracted from mouse embryonic stem cells (R1), which was reverse-transcribed into cDNA. Then PCR product of Oct4 mRNA were transcribed into Oct4 mRNA. Finally the candidate RNA-binding proteins were eluted applying the streptomycin affinity chromatography, and submitted for high-performance liquid chromatography combined with the mass spectrometry. The identified RNA binding proteins were further confirmed by RNA immunoprecipitation (RIP). Results: 121 RNA binding proteins of Oct4 3' untranslated region (UTR) and Oct4 5' UTR were identified with liquid chromatography / mass spectrometry. There were 11 proteins with the number of peptide spectrum matches more or equal to 2, and 7 of them were selected for additional confirmation using RIP method. RIP results showed that Oct4 interacted with DSP, SOD1, LMNA, NPM1, PSIP1, NCL and HDGF. Among them, HDGF had the strongest binding ability to Oct4 mRNA. Conclusion: Identification of Oct4 RNA binding proteins will provide a theoretical basis for the regulation mechanism of Oct4, and will be a basis for the further study of its function.

Effect of apicidin on glioblastoma U87 cells and its regulation of OCT-4 gene expression / 肿瘤研究与临床

Objective@#To investigate the effect of histone deacetylase inhibitor apicidin on the glioblastoma U87 cells and its regulation of OCT-4 gene expression.@*Methods@#Glioblastoma U87 cells were treated with different concentrations of apicidin, and dimethyl sulfoxide instead of apicidin was negative control. Methyl thiazolyl tetrazolium (MTT) assay was used to detect the proliferative ability of U87 cells treated by apicidin. The cell apoptosis was observed under the fluorescence microscope, and the cell cycle was detected by using flow cytometry. Reverse transcription-polymerase chain reaction and Western blot was used to detect the expression of mRNA and protein of U87 cells, respectively relative to the expression of GAPDH.@*Results@#MTT assay results showed that apicidin inhibited U87 cells proliferation in a dose-dependent and time-dependent manner, and half of the inhibitory concentration of cell proliferation at 48 h was (1.74±0.13) μmol/L. The cell proportion of U87 cells in S-phase of the negative control, 0.2, 0.5, and 1.0 μmol/L apicidin was (32.68±0.49)%, (33.73±0.76)%, (42.92±0.56)%, and (56.95±0.53)%, respectively after 48 h apicidin administration (P < 0.05), while the proportion of G1 and G2 phase cells was decreased. The karyopyknosis and other apoptotic changes were detected in U87 cells after 48 h treatment of 1.0 μmol/L apicidin under the confocal fluorescence microscope. Western blot and RT-PCR showed that the mRNA and protein relative levels of U87 cells OCT-4 were reduced after 1.0 μmol/L apicidin treatment for 48 h compared with the negative control group (mRNA: 72.44±0.00 vs. 56.66±0.23; protein: 86.59±0.19 vs. 56.04±0.15, both P < 0.01).@*Conclusions@#Apicidin can inhibit the growth of glioblastoma U87 cells, induce cell cycle arrest and apoptosis. Its mechanism may be related to the expression of OCT-4 inhibited by apicidin.

Doxorubicin induces stemness of mouse TNBC 4T1 cells through Stat3-Oct-4/CD44 signaling pathway / 中国病理生理杂志

AIM:To investigate the stemness of mouse triple-negative breast cancer (TNBC) 4T1 cells induced by doxorubicin (DOX) and the underlying mechanism.METHODS:The 4T1 cells and MDA-MB-468 cells were treated with DOX at different concentrations (0, 0.05, 0.1 and 0.5μmol/L) for 24 h, and the shape and viability of the cells were observed.The concentration of DOX at 0.1μmol/L was chosen as the optimal concentration for the following experiments.The 4T1 cells and MDA-MB-468 cells resistant to DOX were established by continuous stimulation with DOX for 4 weeks, and named as 4T1-DOX and MDA-MB-468-DOX.Sphere formation assay was used to detect the stemness of 4T1cells and MDA-MB-468 cells.The expression of CD133 was observed by immunofluorescence staining.The expression of CD44 was analyzed by flow cytometry.The protein levels of Stat3, phosphorylated Stat3 (p-Stat3) and Oct-4 were determined by Western blot.RESULTS:The sphere formation ability of the 4T1-DOX cells was stronger than that of the 4T1control cells.The 4T1-DOX cells expressed high levels of the stemness markers CD133 and CD44 as compared with the 4 T1 cells (P<0.05).Furthermore, the 4T1-DOX cells exhibited enhanced activation of Stat3 (p-Stat3) and increased expression of Oct-4 (P<0.05) , while the expression of total Stat3 had no obvious variation.In addition, when activation of Stat3 was inhibited by WP1066, the protein levels of p-Stat3, Oct-4 and CD44 were down-regulated (P<0.05).Furthermore, inhibition of Stat3 phosphorylation reduced the sphere formation ability of the 4T1-DOX cells (P<0.05).CONCLUSION:DOX induces the stemness of mouse TNBC 4T1 cells through Stat3-Oct-4 signaling pathway.

Teratoma inmaduro (maligno) de ovario / Ovarian immature (malignant) teratoma

Resumen El teratoma inmaduro se describió por primera vez en 1960 y puede ser puro o estar mezclado con un componente maduro. Es la segunda neoplasia maligna más común de células germinales de todos los cánceres de ovario (representa <1%). Alrededor del 50% de los teratomas inmaduros puros del ovario ocurren en mujeres entre las edades de 10 y 20 años. Se debe considerar el tratamiento para preservar su fertilidad futura porque la mayoría de los tumores de células germinales de ovario son curables con la cirugía conservadora y la quimioterapia combinada de seguimiento. La mayoría de los pacientes diagnosticados con un teratoma inmaduro se curan de su enfermedad. Sin embargo en todas las pacientes, se recomienda un seguimiento cercano, particularmente en los primeros 2 años después del diagnóstico, donde existe una mayor probabilidad de recurrencia.

Abstract The immature teratoma was first described in 1960 and cay be pure or mixed with a mature component. It is the second most common malignant germ cell neoplasm of all ovarian cancers (representing <1%). About 50% of pure immature teratomas of the ovary occur in women between the ages of 10 and 20 years. Treatment should be considered to preserve future fertility because the majority of ovarian germ cell tumors are curable with conservative surgery and combined combination chemotherapy. Most patients diagnosed with an immature teratoma are cured of their disease. However, in all patients, we recommend close follow-up, particularly in the first 2 years after diagnosis, where there is a greater chance of recurrence.

Expressions of OCT-4 and CD44 in laryngeal squamous cell carcinoma and the clinicopathological significance of OCT-4 and CD44 double positive / 肿瘤研究与临床

Objective@#To detect the expression of nuclear transcription factor OCT-4 in laryngeal squamous cell carcinoma (LSCC) and its relationship with CD44, and to explore the relationship between OCT-4 and the cancer stem cells and its significance in tumors.@*Methods@#Sixty-nine specimens of laryngeal surgical resection were selected from LSCC patients from January 2008 to December 2012 in Lanzhou General Hospital of Lanzhou Military Region. Another 18 cases of normal laryngeal mucosa tissues were selected as control. OCT-4, CD44 and OCT-4+ CD44+ cells were observed by using tissue microarray, immunohistochemistry staining and immunohistochemical double staining techniques, and the relationship between their pathological features, such as expression intensity, distribution, cell morphology and clinical parameters were analyzed to explore the clinicopathological significance of CD44+ OCT-4+ cells.@*Results@#The percentage of CD44-positive cells in LSCC was significantly higher than that in normal laryngeal mucosa tissues [(63.62±8.76)% vs. (13.61±5.81)%, P < 0.001]. The percentage of OCT-4-positive cells in LSCC was (50.44±10.84)%, while it was not expressed in normal laryngeal mucosa tissues (P < 0.001). In LSCC, the expression of OCT-4 was significantly lower than that of CD44 (P < 0.001); the coexpression rate of these two molecules was (5.07±2.90)%. The percentage of CD44-positive cells in poorly differentiated LSCC was significantly higher than that in the well-differentiated group [62.78% (8.22%) vs. 28.62% (30.45%), P < 0.05]. The percentage of OCT-4-positive cells in poorly differentiated LSCC was significantly higher than that in the well-differentiated group [49.89% (13.12%) vs. 33.00% (20.00%), P < 0.05]. In the cancer stem cells, the percentage of OCT-4+ CD44+ cells in poorly differentiated carcinoma group was also significantly higher than that in the well-differentiated carcinoma group (P < 0.01), the lower the degree of differentiation, the higher the expression level of OCT-4. In LSCC poorly differentiated group, the expressions of OCT-4 and CD44 showed no significant difference between different T stages, but the percentage of OCT-4+ CD44+ cells elevated with T stage increasing (P < 0.05). The expression of OCT-4 in patients with lymph node metastasis was significantly higher than that in patients without lymph node metastasis (P < 0.01).@*Conclusions@#The expressions of OCT-4 and CD44 are associated with the malignant biological behaviors of LSCC closely. OCT-4 may be a valuable marker of tumor stem cells of LSCC.

Relationship between expressions of OCT-4, CD117 and clinicopathological features and prognosis of patients with ovarian cancer / 肿瘤研究与临床

Objective@#To investigate the relationship between expressions of OCT-4, CD117 and clinicopathological features and prognosis of patients with ovarian cancer.@*Methods@#A total of 70 paraffin-embedded tissues of patients with ovarian cancer from January 2010 to February 2016 in Shanxi Provincial Cancer Hospital were collected. The expressions of OCT-4 and CD117 were detected by immunohistochemistry.@*Results@#OCT-4 was mainly expressed in cytoplasm, while CD117 was expressed in cell membrane and cytoplasm. The positive expression rate of OCT-4 was 74.3% (52/70), and the positive expression rate of CD117 was 68.6% (48/70). The positive expression rates of OCT-4 in ovarian cancer tissues with poorly differentiation and high CA125 levels (≥500 U/ml), no peritoneal effusion and sensitive to chemotherapy drugs were 92.1% (35/38), 87.5% (28/32), 88.9% (24/27), and 78.7% (48/61), respectively, which were higher than those in ovarian cancer tissues with well and moderately differentiation, low CA125 levels (<500 U/ml), peritoneal effusion and resistance to chemotherapy drugs, the differences were statistically significant (P values were 0.000, 0.020, 0.047, and 0.043). The positive expression rates of CD117 in ovarian cancer tissues with poorly differentiation and peritoneal effusion were 84.2% (32/38) and 79.1% (34/43), respectively, which were higher than those in ovarian cancer tissues with well and moderately differentiation and no peritoneal effusion, the differences were statistically significant (P values were 0.006 and 0.017). Patients with OCT-4-positive expression, peritoneal effusion, and poorly differentiation had a shorter overall survival time (all P < 0.05). The peritoneal effusion and differentiation were independent prognostic factors in patients with ovarian cancer (both P < 0.05).@*Conclusion@#OCT-4 can be used as an important reference for predicting drug sensitivity and evaluating prognosis in patients with ovarian cancer.

Inhibitory effect of metformin on aldehyde dehydrogenase positive gastric cancer stem cells and its mechanism / 解放军医学杂志

Objective To investigate the inhibitory effect of metformin (Met) on ALDH positive (ALDH+) gastric cancer stem cells and its mechanism.Methods ALDH+ and ALDH cells were isolated from human gastric cancer cell line MKN45 by flow cytometry.The characteristics of cancer stem cells of ALDH+ cells was verified by self-renewing ability,differentiation capacity experiments and tumorigenicity in nude mice.1mmol/L Met group,5mmol/L Met group and control group (MKN45 cell) were set up.After being acted on MKN45 cell for 48h,the proportion ofALDH+ cells in each group was detected by flow cytometry.RT-PCR test was performed to detect the expression of Oct4,Sox2 and AKT genes in the 3 groups.Results The cell identification showed that the self-renewal ability,differentiation capacity and tumorigenicity of ALDH+ cells were higher than that of ALDH-cells.Drug experiments indicated that the proportion ofALDH+ cells in control group,lmmol/L Met group and 5mmol/L Met group were 36.5％ ± 5.4％,15.6％ ± 1.9％ and 7.6％ ± 1.6％,respectively.The difference between the 3 groups was statistically significant (P＜0.01).RT-PCR revealed that the expressions of Sox2 and AKT genes decreased after Met treatment,and decreased with the increase of Met concentration.The expression of Oct4 gene was higher in 1 mmol/L Met group than in control group,and was lower in 5mmol/L Met group than in control group.Conclusion Met may inhibit the growth ofALDH+ gastric cancer stem cells and the expression ofAKT gene,and can be used as a target drug for the clinical treatment of gastric cancer.

Expression of Oct4 and its relationship with clinicopathological parameters in non-small cell lung cancer / 国际检验医学杂志

Objective To explore the expression of Octamers binding transcription factor 4 (Oct4) and its relationship with clinicopathological parameters in the non-small cell lung cancer (NSCLC) tissues .Methods From September 2013 to December 2016 ,100 cases of NSCLC patients with lung cancer (group NSCLC ,n=100) and their corresponding normal paracancerous tissues (the control group ,n=100) were selected as the samples .The expression of Oct4 mRNA and protein in two groups of tissue samples were detected by fluores-cence quantitative PCR ,immunohistochemistry and western blot .The relationship between the level of Oct4 expression and the clinicopathological parameters of the patients was analyzed ,and the prognosis of the Oct4 positive and negative patients in the NSCLC group was compared .Results The expression of Oct4 mRNA in NSCLC group was (3 .21 ± 0 .18) ,protein expression was (0 .74 ± 0 .11) and the positive rate was 74 .00％ , which were significantly higher than those of control group [(1 .47 ± 0 .11) ,(0 .10 ± 0 .03) ,10 .00％ ,P<0 .05] .The positive rate of Oct4 expression in NSCLC patients was not related to sex ,age ,smoking index and tumor size (P>0 .05) ,but related to pathological type ,degree of differentiation ,and TNM staging ,in which the positive rates of Oct4 in patients with adenocarcinoma ,low differentiation and TNM stage Ⅲ stage were significantly higher than those of non adenocarcinoma ,middle-high differentiation degree ,Ⅰ - Ⅱ TNM stage patients (P< 0 .05) .In NSCLC patients ,compared with Oct4 negative patients ,the distant metastasis rate (64 .86％ ) ,pulse tube infiltration rate (47 .30％ ) in Oct4 positive patients were significantly higher ,and the 1 year survival rate (67 .57％ ) decreased (P<0 .05) .Conclusion The expression of Oct4 in NSCLC tissue is closely related to its clinicopathological parameters .Oct4 is highly expressed in adenocarcinoma ,and with thedecrease of differentiation and progression of tumor stages ,the expression increases and affects the prognosis . Oct4 can be used as a specific diagnostic marker of NSCLC .

Effect of docetaxel on cancer stem cell-like cells in the mucoepidermoid carcinoma cell line MC3 / 医学研究生学报

Objective Docetaxel is commonly used in chemotherapy for patients with mucoepidermoid carcinoma (MEC) in the salivary gland after operation,but it cannot kill all residual tumor cells and prevent recurrence.Cancer stem cells (CSCs) may be responsible for tumor recurrence and drug resistance.Based on the theory of CSCs,the authors investigated the effects of docetaxel on cancer stem cell-like cells in the MEC cell line MC3.Methods MC3 cells in the logarithmic growth phase were divided into a negative control and a docetaxel (40 ng/mL) group.The colony formation rate of the MC3 cells was calculated with the soft agar cloning technique,the growth of the cells detected by MTT assay,the protein and mRNA expressions of CD44 and Oct4 determined by immunocytochemistry and flow cytometry,and tumorigenicity observed by nude mouse tumorigenicity assay.Results Compared with the negative control group,the MC3 cells treated with docetaxel exhibited significant increases in the colony formation rate ([9.14±0.75] vs [33.47±1.30]％,P＜0.05),the protein expressions of CD44 (14.47±0.15 vs 99.50±0.30,P＜0.05) and Oct4 (1.37±0.06 vs 14.60±0.36,P＜0.05),and the mRNA expressions of CD44 (0.207±0.009 vs 0.651±0.015,P＜0.05) and Oct4 (0.223±0.008 vs 2.228±0.005,P＜0.05).At 2 months after injection of 1×103,1× 104,and 1×105 MC3 cells,tumor formation was observed in 0,1,and 3 of the nude mice,respectively,but not in the negative control group.Conclusion MC3 cells surviving docetaxel treatment have the properties of stem cells,and docetaxel can enrich cancer stem cells in the MC3 cells,which plays a key role in tumor recurrence.

Correlation of Cancer Stem-Cell Markers OCT4, SOX2, and NANOG with Clinicopathological Features and Prognosis in Operative Patients with Rectal Cancer

PURPOSE: To investigate the association of cancer stem-cell markers [octamer-binding transcription factor 4 (OCT4), sex determining region Y-box 2 (SOX2), and Nanog homebox (NANOG)] expression with clinicopathological properties and overall survival (OS) in operative rectal cancer (RC) patients receiving adjuvant therapy. MATERIALS AND METHODS: 153 patients with primary RC receiving surgery were enrolled. Tumor tissue and paired adjacent normal tissue sample were collected, and OCT4, SOX2, and NANOG expressions were assessed by immunofluorescent staining. The median follow-up duration was 5.2 years, and the last follow-up date was August 2016. RESULTS: Tumor tissue OCT4 (p < 0.001), SOX2 (p=0.003), and NANOG (p < 0.001) expressions were higher than those in adjacent tissue. OCT4 expression was positively correlated with pathological grade (R=0.185, p=0.022), tumor size (R=0.224, p=0.005), and N stage (R=0.170, p=0.036). NANOG expression was positively associated with tumor size (R=0.169, p=0.036). Kaplan-Meier suggested that OCT4+ was associated with worse OS compared with OCT4− (p < 0.001), while no association of SOX2 (p=0.121) and NANOG expressions (p=0.195) with OS was uncovered. Compared with one or no positive marker, at least two positive markers were associated with shorter OS (p < 0.001), while all three positive markers were correlated with worse OS compared with two or less positive markers (p < 0.001). Multivariate Cox's analysis revealed that OCT4+ (p < 0.001) and N stage (p=0.046) were independent factors for shorter OS. CONCLUSION: Tumor tissue OCT4 expression was correlated with poor differentiation, tumor size, and N stage, and it can serve as an independent prognostic biomarker in operative patients with RC receiving adjuvant therapy.

Expression of Oct4 and its prognostic role in left-and right-side colon cancer / 中国肿瘤临床

Objective:To investigate the expression and prognostic value of the Oct4 protein in colon cancer.Methods:Immunohistochemical technique was used to examine the expression of Oct4 protein in 89 left side colon cancer(LCC)tissues and 77 right-side colon cancer(RCC) tissues.The relationship among Oct4 expression,clinicopathological parameters,and the prognostic value of Oct4 in colon cancer was analyzed.Results:The positive rate of Oct4 protein in LCC was 68.54%,and that in RCC was 71.43%.There was no significant difference between the two values.In addition,Oct4 expression in RCC was positively correlated with histological grade,lymph node metastasis,and Dukes staging.By contrast,Oct4 expression in LCC was only positively correlated with histological grade and Dukes staging.In survival analysis, the 5-year survival rate of LCC was significantly higher than that in RCC(P<0.01).In patients with LCC,no obvious correlation was found between positive and negative Oct4 expression levels in OS.In patients with RCC,Oct4 overexpression indicated poor prognosis(P<0.05). Also,in Cox survival analysis,Oct4 overexpression indicated poor prognosis in RCC but not in LCC.Conclusion:These results indicated that Oct4 plays different roles in LCC and RCC.These roles can be used as theoretical basis for exploring new targets for the diagnosis and treatment of colon cancer.

Overexpression of Oct4B1 induces epithelial mesenchymal transition in colorectal cancer SW480 cells / 中国免疫学杂志

Objective:To investigate whether the overexpression of Oct4B1 gene induces epithelial mesenchymal transition in human colorectal cancer SW480 cells and its possible mechanism.Methods: Experimental group(SW480-Oct4B1):Transfection of SW480 cell lines in colorectal cancer with Oct4B1 overexpression plasmid;Control group(SW480-Oct4B1):negative control plasmid with G418 resistance.Stably transfected cell lines were obtained by G418 culture medium.The two groups were compared with:①Detection of Oct4B1 gene expression in stably transfected cell lines by RT-qPCR;②Scratches and Transwell assays were used to estimate migration and invasion;③Detection of EMT related markers E-cadherin,N-cadherin and Vimentin protein expression by Western blot assay;④Detection of Twist gene and protein expression by RT-PCR and Western blot assays.Results: The transient transfection was confirmed by RT-qPCR and the stable transfected cell lines were obtained from two groups of cells transfected with G418 culture medium.Compared with the control group:①RT-qPCR revealed increased expression of Oct4B1 gene in the experimental group(P<0.01);②Cell migration and invasion were significantly increased(P<0.01);③Epithelial marker:the expression of E-cadherin protein was significantly decreased (P<0.01),interstitial marker:the expression of N-cadherin and Vimentin protein was significantly increased (P<0.01);④Twist mRNA and protein expression were significantly increased(P<0.01).Conclusion: Overexpression of Oct4B1 gene can induce epithelial mesenchymal transition in human colorectal cancer SW480 cells,its molecular mechanism may be related to the promotion of Twist expression.

Expression of Oct4 and its prognostic role in left-and right-side colon cancer / 中国肿瘤临床

Objective:To investigate the expression and prognostic value of the Oct4 protein in colon cancer.Methods:Immunohistochemical technique was used to examine the expression of Oct4 protein in 89 left side colon cancer(LCC)tissues and 77 right-side colon cancer(RCC) tissues.The relationship among Oct4 expression,clinicopathological parameters,and the prognostic value of Oct4 in colon cancer was analyzed.Results:The positive rate of Oct4 protein in LCC was 68.54%,and that in RCC was 71.43%.There was no significant difference between the two values.In addition,Oct4 expression in RCC was positively correlated with histological grade,lymph node metastasis,and Dukes staging.By contrast,Oct4 expression in LCC was only positively correlated with histological grade and Dukes staging.In survival analysis, the 5-year survival rate of LCC was significantly higher than that in RCC(P<0.01).In patients with LCC,no obvious correlation was found between positive and negative Oct4 expression levels in OS.In patients with RCC,Oct4 overexpression indicated poor prognosis(P<0.05). Also,in Cox survival analysis,Oct4 overexpression indicated poor prognosis in RCC but not in LCC.Conclusion:These results indicated that Oct4 plays different roles in LCC and RCC.These roles can be used as theoretical basis for exploring new targets for the diagnosis and treatment of colon cancer.