Immunoexpression of ALDH1A1, FGFR2, CD44 and Caspase-3 in Oral Squamous Cell Carcinoma and Leukoplakia: A pilot study

Current research highlighted the importance to recognize feasible biomarkers for early diagnoses and treatment in oral cancer. Our study analyzed the expression and spatial distribution of ALDH1A1, FGFR2, caspase-3, and CD44 in Oral Squamous Cell Carcinoma (OSCC) and leukoplakia with and without oral mucosal dysplasia. Paraffin-embedded samples of OSCC (n=5), leukoplakia with (n=5) and without (n=5) dysplasia obtained by incisional biopsies were processed using conventional histochemical techniques. Immunohistochemistry was performed using antibodies against ALDH1A1, FGFR2, caspase-3, and CD44. Images of the immunohistochemically stained tissue sections were analyzed according to the intensity of the immunostaining of each marker and classified in Scores. The Kruskal- Wallis test was performed (p≤0.05). Our results demonstrated a statically difference in the expression of all immunomarkers between OSCC and leukoplakia without dysplasia, being more significant in FGFR2 and ALDH1A1. Within the limitations of this study, our data showed that all biomarkers were overexpressed in OSCC and leukoplakia with oral mucosa dysplasia, suggesting that the presence of dysplasia is a significant clinic-pathologic predictor for malignant transformation.

La actual evidencia científica enfatiza la importancia de reconocer biomarcadores viables para el diagnóstico y tratamiento temprano del cáncer oral. Nuestro estudio piloto analizó la expresión y distribución espacial de ALDH1A1, FGFR2, caspasa-3 y CD44 en carcinoma oral de células escamosas (COCE) y en leucoplasia con o sin displasia de la mucosa oral. Las muestras incluidas en parafina de COCE (n=5), con (n=5) y sin (n=5) displasia fueron obtenidas mediante biopsias incisionales, las cuales se procesaron utilizando técnicas histoquímicas convencionales. El análisis inmunohistoquímico se realizó utilizando anticuerpos contra ALDH1A1, FGFR2, caspasa-3 y CD44. Las imágenes de las secciones de cada muestra fueron analizadas según la intensidad de inmunoexpresión de cada marcador y se clasificaron en diferentes escalas (scores). Se realizó la prueba de Kruskal-Wallis (valores de p<0,05). Nuestros resultados demostraron una diferencia estadística en la expresión de todos los inmunomarcadores entre COCE y las muestras con leucoplasia sin displasia, siendo más significativa en FGFR2 y ALDH1A1. Considerando las limitaciones de este estudio, los datos sugieren que la presencia de displasia en la mucosa oral es un importante predictor clínico-patológico de transformación maligna.

Possible role of ALDH1 and CD44 in lip carcinogenesis

Abstract Lip squamous cell carcinoma (LSCC) accounts for 12% of all head and neck cancers. It is caused by chronic exposure to ultraviolet light solar radiation and related to previous actinic cheilitis (AC). This study aimed to investigate the immunostaining of the putative cancer stem cells (CSC) markers ALDH1 and CD44 in AC (n=30) and LSCC (n=20). ALDH1 positivity was found to be statistically higher in LSCC than in AC lesions (p=0.0045), whilst CD44 expression was statistically higher in AC than in LSCC lesions (p=0.0155). ALDH1+ cells in AC lesions were associated with specific clinical features: a younger age (<60 years old), the female gender, white skin, not smoking or consuming alcohol, and a fast evolution, and not associated with the chronic exposure to UV radiation (p<0.0001). CD44 positivity was associated with patients who were male, feoderm, smoked, consumed alcohol, underwent occupational exposure to UV-radiation, and demonstrated lesions with log-time evolution (p<0.0001). ALDH1 + cells were associated with mild dysplasia using a system from the World Health Organization (WHO), and with a low risk of malignant transformation, according to the binary system (p<0.0001). CD44+ cells were also associated with moderated dysplasia, according to the WHO system. In LSCC, ALDH1 + cells were positively associated with patients who were older (≥ 60 years old), smokers, and with those who consumed alcohol (p<0.0001). CD44 + cells in LSCC were associated with older (≥ 60 years old) patients as well, but also with female patients, white skin, non-smokers, and individuals who did not consume alcohol (p<0.0001), all of whom showed distinct patterns in pre- and malignant lesions of both markers. Additionally, in LSCC, both ALDH1 and CD44 staining were associated with smaller tumor sizes (T1/T2; p<0.0001). In summary, although both ALDH1 and CD44 were associated with the presence of dysplasia in AC lesions, the present findings suggest that ALDH1 and CD44 may be activated by different etiopathogenic pathways, predominantly in distinct steps of oral carcinogenesis. CD44 would thus be more significantly related to the potentially malignant lesion, while ALDH1 would be closely linked to malignancy.

Expression Patterns of Inducible Cre Recombinase Driven by Differential Astrocyte-Specific Promoters in Transgenic Mouse Lines / 神经科学通报·英文版

Astrocytes are the most abundant cell type in the central nervous system (CNS). They provide trophic support for neurons, modulate synaptic transmission and plasticity, and contribute to neuronal dysfunction. Many transgenic mouse lines have been generated to obtain astrocyte-specific expression of inducible Cre recombinase for functional studies; however, the expression patterns of inducible Cre recombinase in these lines have not been systematically characterized. We generated a new astrocyte-specific Aldh1l1-CreER knock-in mouse line and compared the expression pattern of Cre recombinase between this and five widely-used transgenic lines (hGfap-CreER from The Jackson Laboratory and The Mutant Mouse Resource and Research Center, Glast-CreER, Cx30-CreER, and Fgfr3-iCreER) by crossing with Ai14 mice, which express tdTomato fluorescence following Cre-mediated recombination. In adult Aldh1l1-CreER:Ai14 transgenic mice, tdTomato was detected throughout the CNS, and five novel morphologically-defined types of astrocyte were described. Among the six evaluated lines, the specificity of Cre-mediated recombination was highest when driven by Aldh1l1 and lowest when driven by hGfap; in the latter mice, co-staining between tdTomato and NeuN was observed in the hippocampus and cortex. Notably, evident leakage was noted in Fgfr3-iCreER mice, and the expression level of tdTomato was low in the thalamus when Cre recombinase expression was driven by Glast and in the capsular part of the central amygdaloid nucleus when driven by Cx30. Furthermore, tdTomato was clearly expressed in peripheral organs in four of the lines. Our results emphasize that the astrocyte-specific CreER transgenic lines used in functional studies should be carefully selected.

Expression and Significance of ALDH1 in Cancer Stem Cell-like Characteristics in Breast Cancer Cell Line MCF-7 / 医学研究杂志

Objective To investigate the expression and significance of ALDH1 in cancer stem cell-like characteristics in breast cancer cell line MCF-7.Methods RNA lenti-virus particles targeting ALDH1 gene were transfected into MCF-7 cells.Subclones with stable ALDH1 expression were selected by puromycin and named MCF-ALDH1 cells.The mRNA and protein expression of ALDH1 were detected by real-time quantitative PCR and Western blot tests.Cell differentiation,stem cell potential were observed by clone formation assay and suspension sphere-forming assay.The abilities of tumor-formation of cells were detected through inoculating the cells into the nod mouse.To detect the drug-resistance ability of cells by MTT test.Results Compared to the MCF-7 cells,the expression of mRNA and protein of ALDH1 by MCF-ALDH1 cells were significantly increased.Clony forming numbers(128 ±22) of the MCF-AL-DH1 cells was higher than controls(46 ± 15)(P ＜ 0.05),The proportion of sphere-initiating cells in MCF-ALDH1 and controls was (16 ± 2) versus(8.0 ± 1.5,P ＜ 0.05).The average tumor volume of MCF-ALDH1 was greater than controls(P ＜ 0.05).The drug resistance index of MCF-ALDH1 was higher than controls.Conclusion Experimental findings from this study showed that MCF-7 cells highly expressing ALDH1 genes,with higher proliferation and differentiation,in vivo tumorigenicity and potential of stem cells.ALDH1 may be used as an target to cure breast cancer.

Novel compound heterozygous mutations of ALDH1A3 contribute to anophthalmia in a non-consanguineous Chinese family

Abstract Anophthalmia is a rare eye development anomaly resulting in absent ocular globes or tissue in the orbit since birth. Here, we investigated a newborn with bilateral anophthalmia in a Chinese family. Exome sequencing revealed that compound heterozygous mutations c.287G > A (p.(Arg96His)) and c.709G > A (p.(Gly237Arg)) of the ALDH1A3 gene were present in the affected newborn. Both mutations were absent in all of the searched databases, including 10,000 in-house Chinese exome sequences, and these mutations were confirmed as having been transmitted from the parents. Comparative amino acid sequence analysis across distantly related species revealed that the residues at positions 96 and 234 were evolutionarily highly conserved. In silico analysis predicted these changes to be damaging, and in vitro expression analysis revealed that the mutated alleles were associated with decreased protein production and impaired tetrameric protein formation. This study firstly reported that compound heterozygous mutations of the ALDH1A3 gene can result in anophthalmia in humans, thus highlighting those heterozygous mutations in ALDH1A3 should be considered for molecular screening in anophthalmia, particularly in cases from families without consanguineous relationships.

Chromatin remodeling protein MORC2 promotes a breast cancer stem-like phenotype by regulating ALDH1A3 expression / 中国癌症杂志

Background and purpose: MORC2 (microrchidia family CW-type zinc finger 2, MORC2) is a newly identified chromatin remodeling protein that plays key roles in DNA-based biological processes including gene transcription and DNA damage repair. However, its functional role in breast cancer development and progression re-mains unknown. ALDH1A3 (aldehyde dehydrogenase 1 family member A3), a member of the aldehyde dehydrogenases (ALDH) superfamily, is a putative breast cancer stem cell marker, but its regulatory mechanism in breast cancer is poor-ly characterized. This study aimed to investigate the effects of knockdown of endogenous MORC2 on the expression levels of ALDH1A3 and the breast cancer stem-like phenotype in MCF-7 cells. Methods: Human breast cancer MCF-7 cells were infected with negative control short hairpin RNAs (shNC) and specific shRNAs targeting human MORC2 (shMORC2), followed by selection with puromycin to generate stable MORC2 gene knockdown cell lines. Western blot and real-time fluorescent quantitative polymerase chain reaction (RTFQ-PCR) were used to examine the protein and mRNA levels of ALDH1A3 in MCF-7 cells stably expressing shNC and shMORC2. Microsphere formation and fluo-rescence-activated cell sorting (FACS) assays were used to analyze the effects of knockdown of MORC2 on the breast cancer stem-like phenotype. Results: Western blot and RTFQ-PCR analyses revealed that the protein and mRNA levels of ALDH1A3 were significantly down-regulated in shMORC2 expressing cells as compared with shNC -transfected control cells. Moreover, mammosphere formation assay showed that knockdown of endogenous MORC2 in MCF-7 cells significantly reduced the ability of cells to form microspheres. Consistently, FACS assays demonstrated that shMORC2-transfected cells had a lower proportion of ALDH-positive stem cells as compared with shNC expressing cells. In contrast, knockdown of MORC2 did not significantly affect the CD44+CD24- stem cell population. Conclusion:MORC2 promotes a breast cancer stem-like phenotype through, at least in part, regulating ALDH1A3 expression.

AAV-Mediated Astrocyte-Specific Gene Expression under Human ALDH1L1 Promoter in Mouse Thalamus

Adeno-associated virus (AAV)-mediated gene delivery has been proposed to be an essential tool of gene therapy for various brain diseases. Among several cell types in the brain, astrocyte has become a promising therapeutic target for brain diseases, as more and more contribution of astrocytes in pathophysiology has been revealed. Until now, genetically targeting astrocytes has been possible by utilizing the glial fibrillary acidic protein (GFAP) promoter. In some brain areas including thalamus, however, the GFAP expression in astrocytes is reported to be low, making it difficult to genetically target astrocytes using GFAP promoter. To study the function of astrocytes in thalamus, which serves as a relay station, there is a great need for identifying an alternative astrocyte-specific promoter in thalamus. Recently, a new astrocyte-specific promoter of ALDH1L1 has been identified. However, it has not been examined in thalamus. Here we developed and characterized an AAV vector expressing Cre recombinase under the human ALDH1L1 promoter, AAV-hALDH1L1-Cre. To test the cell-type specific expression of AAV-hALDH1L1-Cre, AAV virus was injected into several brain regions of Ai14 (RCL-tdTomato) mouse, which reports Cre activity by tdTomato expression. In thalamus, we observed that tdTomato was found mostly in astrocytes (91.71%), with minimal occurrence in neurons (2.67%). In contrast, tdTomato signal was observed in both neurons and astrocytes of the amygdala (neuron: 68.13%, astrocyte: 28.35%) and hippocampus (neuron: 76.25%, astrocyte: 18.00%), which is consistent with the previous report showing neuronal gene expression under rat ALDH1L1 promoter. Unexpectedly, tdTomato was found mostly in neurons (91.98%) with minimal occurrence in astrocytes (6.66%) of the medial prefrontal cortex. In conclusion, hALDH1L1 promoter shows astrocyte-specificity in thalamus and may prove to be useful for targeting thalamic astrocytes in mouse.

Influence of RhoC on Apoptosis and Cancer-stem-cell-marker-ALDH1A1 and Morphology in Hep2 Laryngeal Squamous Carcinoma Cell / 医学研究杂志

Objective To investigate the effects of RhoC on biological behavior in the laryngeal squamous carcinoma.Methods By delivering exogenous gene into Hep2 laryngeal squamous carcinoma cell line,we alternatively repressed and strengthened the expression of RhoC.We tested the apoptosis of Hep2 tumor cell line with TUNEL,visualized tumor cells shape by staining cell skeleton with Alexa fluor phalloidin,measured the mRNA of CSC marker ALDH1A1 with QPCR.Results After repressing the expression of RhoC in Hep2 cell line,the apoptosis of cancer cells was elevated,the expression of CSC marker ALDH1A1 was significantly decreased.RhoC impacted the shapes of Hep2.Conclusion RhoC havd a positive role in LSCC metastasis.RhoC is a promising target of anti-metastases in LSC.

Research of HIF-1α,ALDH1 and Hedgehog signaling pathway cooperation involved in activation of cancer stem cell in triple negative breast cancer / 中国免疫学杂志

Objective:To explore the cooperation and clinical significance of HIF-1α,ALDH1 and Hedgehog signaling pathway in the activation of cancer stem cell(CSC) in triple negative breast cancer(TNBC).Methods: ALDH1+(Aldehyde dehydrogenase1)breast cancer stem cells and ALDH1-breast cancer cells were selected from MDA-MB-231 cells by magnetic activated cell sorting system(MACS),qRT-PCR method was employed to analyze the expression differences of HIF-1α and Hedgehog signaling molecules Sonic hedgehog(SHH),patched1(PTCH1),Smoothened(SMO) and Glioma-associated oncogene homoglog1(GLI1) in ALDH1+ breast cancer stem cells and ALDH1-breast cancer cells.Immunohistochemical method was applied to study the expressions of HIF-1α and ALDH1 and the relationships among HIF-1α,ALDH1 and Hedgehog signaling molecules in TNBC.Results: The expressions of HIF-1α mRNA,SMO mRNA and GLI1 mRNA in ALDH1+ breast cancer stem cell were higher than those in ALDH1-breast cancer cell(P all<0.05).The positive expression rates of HIF-1α were 90.0% and 70.0%,and the positive rates of ALDH1 were 93.3 % and 66.7 % in TNBC and non-TNBC,respectively(P all<0.05).Spearman rank correlation analysis showed that the expression of HIF-1α was positively related with that of ALDH1 in TNBC(r=0.53,P<0.01).HIF-1α expression was correlated with lymph node metastasis and TNM stage(P all<0.05),ALDH1 expression was correlated with histological grade and TNM stage(P all<0.05).In addition,the expression of HIF-1α was positively related with that of Hedgehog signaling molecules SHH(r=0.584,P<0.01),SMO(r=0.467,P<0.01) and GLI1(r=0.439,P<0.05),the expression of ALDH1 was positively related with that of SHH(r=0.426,P<0.05) and GLI1(r=0.394,P<0.05).Conclusion: HIF-1α and Hedgehog signaling pathway were activated in ALDH1+ breast cancer stem cell.HIF-1α,ALDH1 and Hedgehog molecules may cooperate with each other to activate breast CSC to promote the malignant progression of TNBC.

Expression and significance of ABCG2 and ALDH1 in malignant transformation of ovarian endometriosis / 重庆医学

Objective To investigate the expression and significance of ABCG2 and ALDH1 in malignant transformation of ovarian endometriosis.Methods The protein expressions of ABCG2 and ALDH1 were detected with immunohistochemistry in 38 cases of malignant transformation of ovarian endometriosis(EMs malignant transformation group),35 cases of endometriosis(EMs group)and 30 cases of normal endometrium(control group).Results The positive rates of ABCG2 and ALDH1 expressions in EMs malignant transformation group were significantly higher than those in EMs group and control group(P0.05).There was no correlation between the expression of ABCG2 and ALDH1 in malignant transformation of ovarian endometriosis(P>0.05).Conclusion The expressions of ABCG2 and ALDH1 might be involved in malignant transformation of ovarian endometriosis.

Serum CA125 and ALDH1 levels for diagnosis and postoperative follow-up of endometriosis / 西安交通大学学报(医学版)

ABSTRACT:Objective To investigate CA125 and ALDH1 levels in diagnosis and postoperative follow‐up of endometriosis (EMs) .Methods Serum CA125 and ALDH1 levels were tested by enzyme‐linked immunosorbent assay (ELISA) in 105 patients with EMs and 100 healthy subjects who came to our hospital between January 2011 and June 2013 .Results The preoperative serum CA125 and ALDH1 levels of EMs patients were significantly higher than those in the control group ( P0 .05) .Taking CA125≥35 U/mL and ALDH1≥2 .0 pg/mL as the boundary values ,the specificity , sensitivity ,positive predictive value and negative predictive value of serum CA 125 combined with ALDH1 for diagnosis of EMs were 87 .1% ,91 .6% ,84 .6% and 89 .3% ,respectively .One month after operation ,serum CA125 and ALDH1 levels of EMs patients were decreased significantly (P0 .05) .Relapse occurred in 1 patient 1 year after treatment and in 6 patients 2 years after treatment ;all these patients had a higher serum ALDH1 level than the control group ,but only 2 of them had a higher serum CA125 level .Conclusion The combined detection of serum CA125 and ALDH1 has a higher sensitivity in diagnosis of EMs .Serum ALDH1 level increases in stages Ⅰ and Ⅱ and recurrent patients and can become a better serum marker for early diagnosis and detection of the recurrence of EMs in patients .

The biological characteristics of adriamycin-resistant breast cancer cell line / 肿瘤

Objective: To investigate biological characteristics of adriamycin (ADR)-resistant breast cancer cell line. Methods: Using MCF-7 cells as parent cell line, an ADR-resistant cell line named MCF-7/ADR was established. The half maximal inhibitory concentration (IC50) values of MCF-7/ADR and MCF-7 cells were detected by resazurin test. The ability of colony formation of MCF-7/ADR and MCF-7 cells was measured by colony formation assay. The percentages of CD44+/CD24- and acetaldehyde dehydrogenase 1 (ALDH1)+ subtypes of cancer stem cells in MCF-7/ADR and MCF-7 cells were detected by FCM and immunofluorescent assay, respectively. The expressions of ALDH1 and smoothened (Smo) and Gli1 proteins in Hedgehog signaling pathway related to modulation of stem cells in MCF-7/ADR and MCF-7 cells were detected by Western blotting. Results: The IC50 value of ADR in MCF-7/ADR cells was higher than that in MCF-7 cells (P < 0.01), the resistant fold was 385. The colony formation rate of MCF-7/ADR was higher than that of MCF-7 cells (P < 0.01). The percentages of CD44+/CD24- and ALDH1+ subtypes of cancer stem cells in MCF-7/ADR cells were higher than those in MCF-7 cells (both P<0.01). The expression levels of ALDH1 protein and the Smo and Gli1 proteins in Hedgehog pathway related to modulation of stem cells in MCF-7/ADR cells were higher than those in MCF-7 cells (P < 0.01, P<0.01 and P<0.05, respectively). Conclusion: The percentages of CD44+/CD24- and ALDH1+ subtypes of cancer stem cells in MCF-7/ADR cells are higher, and the expressions of ALDH1 protein and the Smo and Gli1 proteins in Hedgehog pathway related to modulation of stem cells in MCF-7/ADR cells are higher. This mechanism may be responsible for ADR resistance in breast cancer cells.

Isolation and culture of stem cells derived from dental pulp of permanent teeth: a successful method / Isolation and culture of stem cells derived from dental pulp of permanent teeth: a successful method

Objective: To establish cultures of cells from the pulp of permanent teeth by the explant method assessing parameters usually presented by stem cells, such as the expression of certain markers and the differentiation ability into osteogenic, adipogenic, and chondrogenic lineages. This study also aimed to assess the expression of ALDH1 (aldehyde dehydrogenase 1) enzyme activity on the isolated cells. Materials and method: The pulp tissue, obtained from wisdom teeth, was placed in a 6-well plate containing proper culture medium, and stored at 37 °C and 5% CO2 for cell proliferation and plastic adherence. Cells were tested for the expression of surface markers and for ALDH1 enzyme activity, by flow cytometry. In addition, cells were assessedfor multi-differentiation potential. Results: The isolated cells showed high expression of CD44 (98.8%), CD73 (100%), and CD90 (97.2%), and moderate expression of STRO-1 (18.4%) and ALDH1 (16.2%), by flow cytometry. Similarly, the cells showed differentiation ability into all three lineages of cells tested. Conclusion: Our results suggest that the explant method - or cell proliferation method - is suitable for the isolation and cultureof stem cells from dental pulp of permanent teeth. The isolated cells may be considered stem cells, based on the current criteria for their characterization, such as plastic adherence, expression of certain markers, and the absence of others, as well as multi-differentiation potential, which showed to be promising for the application in tissue regeneration.

Effect of low-dose metronomic chemotherapy on ALDH1 expression in human ovarian carcinoma xenograft tumor in nude mice / 肿瘤

Objective: To investigate the influence of maximum-tolerated dose chemotherapy with cisplatin (MTD-DDP) and low-dose metronomic chemotherapy with DDP (LDM-DDP) on human ovarian carcinoma xenograft tumor in nude mice and the expression of aldehyde dehydrogenase 1 (ALDH1) in xenograft tumor. Methods: Murine subcutaneous xenograft tumors of human epithelial ovarian cancer SKOV3 cells were established in nude mice and treated with MTD-DDP and LDM-DDP, respectively. The growth of xenograft tumor was observed. The xenograft tumor primary cells were established. ALDH1-positive cells were obtained by fluorescence-activated cell sorting. The abilities of colony formation and tumorigenicity of ALDH1-positive cells in vitro and in vivo were detected by colony formation assay and tumor formation in nude mice, respectively. The expression levels of Ki-67, breast cancer resistant protein (BCRP) and CD133 in ALDH1-positive cells were detected by Western blotting. Results: The growth of xenograft tumor in nude mice was inhibited by MTD-DDP and LDMDDP, and this inhibition effect was significant in LDM-DDP group (P < 0.001). The proportion of ALDH1-positive cells in xenograft tumor primary cells was lower in MTD-DDP group and higher in LDM-DDP group as compared with that in the control group (0.9% NaCl solution) (both P < 0.001). The abilities of colony formation and tumorigenicity of ALDH1-positive cells were higher than those of ALDH1-negative cells (both P < 0.001). The expression level of Ki-67 was lower but the expression levels of BCRP and CD133 were higher in ALDH1-positive cells than those in ALDH1-negative cells (all P < 0.001). Conclusion: LDM-DDP can effectively inhibit the growth of human ovarian carcinoma xenograft tumors in nude mice and reduce the proportion of ALDH1-positive cells.

Liver cancer stem cells are selectively enriched by low-dose cisplatin

Accumulating evidence has indicated the importance of cancer stem cells in carcinogenesis. The goal of the present study was to determine the effect of low-dose cisplatin on enriched liver cancer stem cells (LCSCs). Human hepatoblastoma HepG2 cells were treated with concentrations of cisplatin ranging from 1 to 5 μg/mL. Cell survival and proliferation were evaluated using a tetrazolium dye (MTT) assay. LCSCs were identified using specific markers, namely aldehyde dehydrogenase-1 (ALDH1) and CD133. The percentage of ALDH1+ or CD133+ cells was examined by flow cytometric analysis. The expression of ALDH1 and/or CD133 in HepG2 cells was determined by immunocytochemical analysis. Low-dose cisplatin treatment significantly decreased cell survival in HepG2 cells after 24 or 72 h. However, the percentage of LCSCs in the surviving cells was greatly increased. The percentage of ALDH1+ or CD133+ cells was increased in a time- and dose-dependent manner after treatment with 1-4 μg/mL cisplatin, whereas 5 μg/mL cisplatin exposure slightly reduced the number of positive cells. These findings indicate that low-dose cisplatin treatment may efficiently enrich the LCSC population in HepG2 cells.

Avaliação da imunomarcação de células-tronco tumorais em carcinossarcomas mamários e carcinomas em tumores mistos em cadelas / Evaluation of immunostaining of tumor stem cells in mammary carcinosarcomas and carcinomas in mixed tumors in bitches

As células-tronco tumorais (CTTs) pertencem a uma pequena população de células dentro do tumor com propriedades de autorrenovação e diferenciação em outros tipos celulares. Neste estudo avaliou-se o comportamento tanto das porções mesenquimais quanto das epiteliais de seis carcinossarcomas (CSs), 11 carcinomas em tumores mistos (CTMs) grau I, 11 grau II e 10 grau III. Nas porções epiteliais dos CS e CTM foram observadas imunomarcações para os anticorpos CD44, CD24, Oct-4 e ALDH-1. Nas porções mesenquimais dos CS, nas porções epiteliais dos CTMs graus II e III não houve imunomarcação para o ALDH-1. Concluiu-se que as CTTs são expressas em proporções iguais tanto nas porções mesenquimais quanto nas epiteliais dos CSs e ausentes nas porções mesenquimais bem diferenciadas de CTMs.

Cancer stem cells belong to a small population of cells within the tumor with properties of self-renewal and differentiation into other cell types. In this study, the behavior of both portions, mesenchymal and epithelial, was evaluated. Six carcinosarcomas (CSs), 11 carcinomas within mixed tumors (CWMTs) grade I, 11 grade II, and 10 grade III were evaluated. In the epithelial portions of the CS and CWMTs was observed immunostaining for antibodies CD44, CD24, Oct-4 and ALDH-1. In the mesenchymal portions of the CS, in the epithelial portions of CMTs grades II and III no immunostaining for ALDH-1 was found. It was concluded that the tumor stem cells are expressed in equal proportions in the epithelial and mesenchymal portions of the CS. No immunostaining in the mesenchymal portions of well-differentiated CWMTs was seen.

Gene Expression Profiles in Genetically Different Mice Infected with Toxoplasma gondii: ALDH1A2, BEX2, EGR2, CCL3 and PLAU

Toxoplasma gondii can modulate host cell gene expression; however, determining gene expression levels in intermediate hosts after T. gondii infection is not known much. We selected 5 genes (ALDH1A2, BEX2, CCL3, EGR2 and PLAU) and compared the mRNA expression levels in the spleen, liver, lung and small intestine of genetically different mice infected with T. gondii. ALDH1A2 mRNA expressions of both mouse strains were markedly increased at day 1-4 postinfection (PI) and then decreased, and its expressions in the spleen and lung were significantly higher in C57BL/6 mice than those of BALB/c mice. BEX2 and CCR3 mRNA expressions of both mouse strains were significantly increased from day 7 PI and peaked at day 15-30 PI (P<0.05), especially high in the spleen liver or small intestine of C57BL/6 mice. EGR2 and PLAU mRNA expressions of both mouse strains were significantly increased after infection, especially high in the spleen and liver. However, their expression patterns were varied depending on the tissue and mouse strain. Taken together, T. gondii-susceptible C57BL/6 mice expressed higher levels of these 5 genes than did T. gondii-resistant BALB/c mice, particularly in the spleen and liver. And ALDH1A2 and PLAU expressions were increased acutely, whereas BEX2, CCL3 and EGR2 expressions were increased lately. Thus, these demonstrate that host genetic factors exert a strong impact on the expression of these 5 genes and their expression patterns were varied depending on the gene or tissue.