OBJECTIVEThis study aims to investigate the expression and relationship of CD44 and CD133 in normal oral mucosa, oral potentially malignant disorder (OPMD), and oral squamous cell carcinoma (OSCC). This work also analyzes the relationship between such expression and clinical factors. This study intends to evaluate the clinical value of using CD44 and CD133 as indices to evaluate the carcinogenic potential of OPMD.

METHODSClinical data from 60 patients with OPMD, 60 patients with OSCC, and 10 cases of normal oral mucosa were analyzed. Double immunohistochemical analysis was applied to investigate the expression of CD44 and CD133 in paraffin sections of normal oral mucosa, OPMD, and OSCC tissues. Subsequently, the relationships between such expression and clinical factors were analyzed.

RESULTSThe positive rates of CD44 expression in the normal oral mucosa, OPMD, and OSCC tissues were 100.00%, 96.67%, and 71.67% (P<0.05), respectively. Meanwhile, the positive rates of CD133 expression in the normal oral mucosa, OPMD, and OSCC tissues were 0.00%, 35.00%, and 63.33% (P<0.05), respectively. The expression of CD44 and CD133 was found to be correlated (P<0.05). Such expression was related to the clinical stages and lymphatic metastasis of OSCC (P<0.05).

CONCLUSIONSCD44 and CD133 can be used individually as clinical indices to evaluate the carcinogenic potential of OPMD.
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AC133 Antigen ; Carcinogenesis ; Carcinoma, Squamous Cell ; Humans ; Hyaluronan Receptors ; Lymphatic Metastasis ; Mouth Mucosa ; Mouth Neoplasms

OBJECTIVETo study the expansion potential of megakaryocyte progenitor cells (MPC) from human placenta tissue CD133+ (PT-CD133+) cells.

METHODSPT-CD133+ cells were purified from mononuclear cells (MNC) by magnetic activated cell sorting (MACS) and seeded in serum-free liquid culture medium supplemented with thrombopoietin (TPO), interleukin-3 (IL-3), and stem cell factor (SCF) to expand MPC. At day 7, 10 and 14, the total cell number was counted and the dynamic changes of CD133, CD34, and CD41 antigens expression during ex-vivo expansion were analyzed by flow cytometry (FCM). PT-CD133+ cells at different expansion time were collected and cultured in collagen semisolid medium for colony forming units-megakaryocyte (CFU-MK) assay.

RESULTSPT-CD133+ cells could be optimally expanded at day 7 by 13 +/- 2 fold increase in serum-free liquid culture systems and the total cell number was expanded by 160 fold at day 14. With the expansion time going on, the expression of CD133, CD34 decreased and that of CD41 increased. The expanded megakaryocytes were immature and no sign of platelet formation. Both PT-CD133+ cells before and after expansion could form CFU-MK, the total number of CFU-MK peaked at day 10 of expansion by 54 +/- 10 fold increase.

CONCLUSIONHuman PT-CD133+ cells have a high capacity of MPC expansion, 10 days culture could give rise to the maximum number of CFU-MK.

AC133 Antigen ; Antigens, CD ; Cell Differentiation ; Cells, Cultured ; Female ; Glycoproteins ; Humans ; Megakaryocyte Progenitor Cells ; cytology ; Peptides ; Placenta ; cytology ; Pregnancy

OBJECTIVETo review the leading roles of glioma stem cells (GSCs) and their sophisticated interactions with other cells in the tissue remodeling process of gliomagenesis.

DATA SOURCESPublished articles about assessing GSCs in tumor initiation, progression, and multiple interactions with other cells in the special microenvironment were selected using PubMed. The search terms were "glioma stem cells", "tumorigenesis", and "microenvironment".

STUDY SELECTIONArticles regarding the tissue remodeling of GSCs in gliomagenesis were selected.

RESULTSGSCs exhibit enhanced tumor-initiating ability, could reestablish tumor, and were resistant to radiotherapy and chemotherapy. Studying the role of GSCs in gliomagenesis helps to develop targeting therapy against GSCs, which seems to be a cure for gliomas. However, sophisticated interactions between GSCs and their local microenvironment during tumor remodeling, including integrating with partially differentiated tumor cells, GSCs niche, neural stem cells (NSCs), normal glia, tumor-infiltrating lymphocytes, may obscure the leading role of GSCs during gliomagenesis, and make single targeting therapy unsuccessful.

CONCLUSIONSUnderstanding the biological behaviour of GSCs and their regulatory mechanisms may directly impact current efforts for more directed therapeutics against the highly aggressive gliomas. For multiple possible sources to turning into GSCs, simply eradicating the existing GSCs is not enough to be a cure for gliomas, blocking the potential sources of GSCs and ameliorating the local tumor inducing/promoting microenvironment should be a reasonable strategy.

AC133 Antigen ; Antigens, CD ; Biomarkers, Tumor ; metabolism ; Glioma ; metabolism ; pathology ; Glycoproteins ; Humans ; Neoplastic Stem Cells ; metabolism ; pathology ; Peptides

OBJECTIVETo determine the association of CD133 expression with the sensitivity to radiotherapy among rectal cancer patients.

METHODSThe clinical data of 32 rectal cancer patients was retrospectively collected for patients who received a short-term preoperative radiotherapy(5 Gy/d,×5 d) from 2008 to 2010. Pretreatment tumor biopsies were immunostained for CD133 expression. Rectal cancer regression grade (RCRG) was used to evaluate the sensitivity of the rectal cancer to preoperative radiotherapy. The correlation of CD133 expression and sensitivity to radiotherapy was analyzed.

RESULTSCD133 differentially expressed in rectal cancer tissue with 17 high expression and 15 low expression. The expression of CD133 was associated with the differentiation of rectal cancer with higher expression of CD133 among poorly differentiated rectal cancers(P<0.05). Among the CD133-high patients, two patients showed 1st RCRG, five patients showed 2nd RCRG and ten patients showed 3rd RCRG. For the CD133-low patients, there were five 1st RCRG, seven 2nd RCRG and three 3rd RCRG. There was a significant association between CD133 expression and sensitivity to radiotherapy (P=0.037). Multivariate logistic regression analysis showed that the expression level of CD133(P=0.027) and the differentiation of rectal cancer(P=0.046) were independent predictive factors for the sensitivity of rectal cancer to radiotherapy.

CONCLUSIONSCorrelation between CD133 expression and sensitivity to radiotherapy of rectal cancer may exist, which may be helpful in predicting the sensitivity of rectal cancer to preoperative radiotherapy.

AC133 Antigen ; Antigens, CD ; metabolism ; Biomarkers, Tumor ; metabolism ; Biopsy ; Combined Modality Therapy ; Glycoproteins ; metabolism ; Humans ; Peptides ; metabolism ; Rectal Neoplasms ; metabolism ; radiotherapy ; Retrospective Studies

OBJECTIVETo study the expressions of Integrinalpha2beta1 and CD133 in benign prostatic hyperplasia (BPH) complicated by prostatitis and their significance.

METHODSSpecimens were obtained from 56 BPH patients undergoing transvesical prostatectomy. Paraffin sections of the specimens were subjected to HE staining for pathological examination of inflammatory changes under the light microscope. Twenty-four patients with simple BPH were included in Group A, and the other 32 with BPH complicated with prostatitis in Group B. The expressions of Integrinalpha2beta1 and CD133 in the prostatic tissues of the two groups were determined by immunohistochemistry, Western blotting and IPP6.0 image analysis software.

RESULTSThe expressions of Integrinalpha2beta1 and CD133 were significantly higher in Group B than in A (P < 0.05), and so were the mean relative value of the optical density of Integrinalpha2beta1 (0.29 +/- 0.18 vs 0.04 +/- 0.03) and that of CD133 (0.08 +/- 0.07 vs 0.0020 +/- 0.0018) (P < 0.05).

CONCLUSIONInflammation can up-regulate the expressions of Integrinalpha2beta1 and CD133 in BPH tissue.

AC133 Antigen ; Antigens, CD ; metabolism ; Glycoproteins ; metabolism ; Humans ; Inflammation ; metabolism ; Integrin alpha2beta1 ; metabolism ; Male ; Peptides ; metabolism ; Prostatic Hyperplasia ; complications ; metabolism ; pathology ; Prostatitis ; complications ; metabolism ; pathology

The present study was aimed to explore the role of c-Myc gene regulation in maintaining the self-renewal and drug-resistant properties of colon cancer stem cells (CSCs) and the underlying mechanism. CD133(+) cells were isolated by flow cytometry cell sorting from human HT29 cancer cells. A small interfering RNA (siRNA) against c-Myc was used, and the mRNA and protein expressions of c-Myc were investigated by real-time PCR and Western blotting, respectively. To evaluate the effect of c-Myc on the drug resistance of colon CSCs, CD133(+) cells transfected with c-Myc-siRNA were exposed to 5-FU, oxaliplatin, or their combination. The expressions of ATP-binding cassette (ABC) transporters, including ABCG2, ABCB5 and MDR-1, were detected by Western blotting. The results showed that c-Myc was highly expressed in CD133(+) colon CSCs, and the protein and mRNA expressions of c-Myc were effectively blocked by c-Myc siRNA. Furthermore, CD133(+) cells showed significantly increased survival rate in chemotherapy treatment, compared with CD133(-) cells. c-Myc silencing sensitized CD133(+) cells to chemotherapy-induced cytotoxicity and down-regulated the protein expression levels of ABCG2, MDR-1 and ABCB5. These results suggest c-Myc silencing may regulate the expressions of ABC transporters in colon CSCs, and enhance the sensitivity of CSCs to the chemotherapy.
AC133 Antigen ; ATP-Binding Cassette Transporters ; Cell Line, Tumor ; Colon ; Down-Regulation ; Humans ; Neoplastic Stem Cells ; Proto-Oncogene Proteins c-myc ; RNA, Small Interfering

OBJECTIVETo investigate the expression of CD133 in the bone marrow of patients with myelodysplastic syndrome (MDS) and explore its clinical significance.

METHODSThe expression of CD133 and CD34/CD38 in the bone marrow was detected using flow cytometry in 31 cases of refractory anemia with excess blasts (RAEB), 10 cases of refractory cytopenia with multilineage dysplasia (RCMD) and 11 cases of aplastic anemia (AA).

RESULTSThe percentage of CD133-expressing cells was 6.75% in patients with RAEB, significantly higher than that in patients with RCMD (1.41%) and AA (2.70%) (P<0.05); the percentage of CD133-positive cells were similar between the latter two patient groups (P>0.05). The percentage of CD34(+)/CD38- cells was similar in the 3 groups (P>0.05), all lower than 1%.

CONCLUSIONSAdvanced MDS patients are characterized by an increase of CD133-expressing cells, suggesting the value of CD133 in the diagnosis of RAEB. CD34(+)/CD38- cells do not show a significant value in the diagnosis of MDS.

AC133 Antigen ; Anemia, Aplastic ; metabolism ; Antigens, CD ; metabolism ; Antigens, CD34 ; metabolism ; Female ; Flow Cytometry ; Glycoproteins ; metabolism ; Humans ; Male ; Middle Aged ; Myelodysplastic Syndromes ; diagnosis ; metabolism ; Peptides ; metabolism

OBJECTIVETo select the peptides that specifically bind human cancer stem cell surface marker CD133 from the Ph.D.-7>(TM) phage peptide library.

METHODSWith a biotinylated extracellular fragment of human cancer stem cell surface marker CD133 as the target protein, the CD133 high-affinity peptides were screened from the phage peptide library by liquid phase panning. The clones with high-binding force with human CD133 were then identified by sandwich ELISA and their single-stranded DNA was extracted to test the specificity by competitive ELISA. The amino acid sequences of the selected peptides derived from the phage DNA sequences were synthesized after sequence alignment analysis, and their capacity of binding with colorectal carcinoma cells was assessed by immunofluorescence technique.

RESULTSAfter 4 rounds of liquid phase selection, the phages capable of specific binding with human CD133 were effectively enriched, with an enrichment ratio of 388 times compared to that at the fourth and first rounds. Thirteen out of the 20 clones from the fourth round of panning were identified as positive clones, among which 11 had identical amino acid sequence of TISWPPR, and 2 had the sequence of STTKLAL, and the former sequence showed a stronger binding specificity to CD133.

CONCLUSIONWe have successfully obtained a peptide that specifically binds human CD133 from the Ph.D.-7(TM) phage peptide library, demonstrating the feasibility of screening small molecule high-affinity polypeptides from phage peptide library by liquid-phase panning.

AC133 Antigen ; Antigens, CD ; metabolism ; Biomarkers, Tumor ; metabolism ; DNA, Single-Stranded ; Glycoproteins ; metabolism ; Humans ; Neoplastic Stem Cells ; metabolism ; Peptide Library ; Peptides ; metabolism ; Protein Binding ; Sequence Analysis, DNA

OBJECTIVETo construct a replication-incompetent adenovirus vector targeting cancer stem cells by modified touchdown PCR and overlap extension PCR and investigate its infection efficiency in CD133(+) SW480 cells in vitro.

METHODSThe two portions of the fiber gene encoding the Ad5 fiber knob domain with the HI loop deleted were amplified using two pairs of designed primers and then linked by overlap extension PCR. The product obtained was identified by sequencing and inserted into prokaryotic expression vector pEGFP-N1. The product, pEGFP-N1 KNOBδHI, contained a unique EcoRV restriction site in the deleted portion of the sequence encoding the HI loop. The gene sequences of the adenovirus fiber were amplified using both common PCR and overlap extension PCR, then identified by sequencing and inserted into pNEB193, resulting in pNEB-F5. CD133(+) SW480 cells were infected with the generated adenovirus vectors Ad5-GFP and Ad5FHI-GFP to investigate the infection efficiency using fluorescent microscope.

RESULTSThe target fragments of expected sizes were amplified by touchdown PCR and overlap extension PCR, but not by common PCR. Ad5FHI-GFP showed a higher infection efficiency than Ad5-GFP in CD133(+) SW480 cells.

CONCLUSIONCompared with common PCR, touchdown PCR and overlap extension PCR can significantly improve the specificity and efficiency of the PCR products for constructing CD133(+) cancer stem cell-selective adenovirus type 5 vector, which provides carriers for tumor-targeted gene therapy.

AC133 Antigen ; Adenoviridae ; genetics ; Antigens, CD ; Cell Line, Tumor ; Genetic Vectors ; Glycoproteins ; Humans ; Neoplastic Stem Cells ; cytology ; virology ; Peptides ; Plasmids ; genetics ; Polymerase Chain Reaction ; methods

BACKGROUNDRecent studies have suggested that cancer stem cells are one of the major causes for tumor recurrence due to their resistance to radiotherapy and chemotherapy. Although the highly invasive nature of glioblastoma (GBM) cells is also implicated in the failure of current therapies, it is not clear how glioma stem cells (GSCs) are involved in invasiveness. Rac1 activity is necessary for inducing reorganization of actin cytoskeleton and cell movement. In this study, we aimed to investigate the distribution characteristics of CD133+ cells and Rac1+ cells in GBM as well as Rac1 activity in CD133+ GBM cells, and analyze the migration and invasion potential of these cells.

METHODSA series of 21 patients with GBM were admitted consecutively and received tumor resection in Tianjin Medical University General Hospital during the first half of the year 2011. Tissue specimens were collected both from the peripheral and the central parts for each tumor under magnetic resonance imaging (MRI) navigation guidance. Immunohistochemical staining was used to detect the CD133+ cells and Rac1+ cells distribution in GBM specimens. Double-labeling immunofluorescence was further used to analyze CD133 and Rac1 co-expression and the relationship between CD133+ cells distribution and Rac1 expression. Serum-free medium culture and magnetic sorting were used to isolate CD133+ cells from U87 cell line. Rac1 activation assay was conducted to assess the activation of Rac1 in CD133+ and CD133 - U87 cells. The migration and invasive ability of CD133+ and CD133 - U87 cells were determined by cell migration and invasion assays in vitro. Student's t-test and one-way analysis of variance (ANOVA) test were used to determine statistical significance in this study.

RESULTSIn the central parts of GBMs, CD133+ cells were found to cluster around necrosis and occasionally cluster around the vessels under the microscope by immunohistological staining. In the peripheral parts of the tumors, CD133+ cells were lined up along the basement membrane of the vessels and myelinated nerve fibers. Rac1 expression was high and diffused in the central parts of the GBMs, and the Rac1+ cells were distributed basically in accordance with CD133+ cells both in the central and peripheral parts of GBMs. In double-labeling immunofluorescence, Rac1 was expressed in (83.14 ± 4.23)% of CD133+ cells, and CD133 and Rac1 co-expressed cells were located around the vessels in GBMs. Significantly higher amounts of Rac1-GTP were expressed in the CD133+ cells (0.378 ± 0.007), compared to CD133- cells (0.195 ± 0.004) (t = 27.81; P < 0.05). CD133+ cells had stronger ability to migrate (74.34 ± 2.40 vs. 38.72 ± 2.60, t = 42.71, P < 0.005) and invade (52.00 ± 2.28 vs. 31.26 ± 1.82, t = 30.76, P < 0.005), compared to their counterpart CD133- cells in transwell cell migration/invasion assay.

CONCLUSIONSThese data suggest that CD133+ GBM cells highly express Rac1 and have greater potential to migrate and invade through activated Rac1-GTP. The accordance of distribution between Rac1+ cells and CD133+ cells in GBMs implies that Rac1 might be an inhibited target to prevent invasion and migration and to avoid malignant glioma recurrence.

AC133 Antigen ; Antigens, CD ; metabolism ; Cell Line, Tumor ; Glioblastoma ; metabolism ; pathology ; Glioma ; metabolism ; pathology ; Glycoproteins ; metabolism ; Humans ; Immunohistochemistry ; In Vitro Techniques ; Peptides ; metabolism ; rac1 GTP-Binding Protein ; metabolism