To investigate the expression of hypoxia-inducible factor 1α (HIF-1α) and CD133 in predicting pathologic remission and survival of patients with locally advanced rectal cancer undergoing neoadjuvant chemoradiotherapy.One hundred and fourteen patients with locally advanced rectal cancer undergoing neoadjuvant chemoradiotherapy from January 2010 to December 2015 in Jinhua Municipal Central Hospital were enrolled in the study. RT-PCR and immunohistochemistry methods were used to detect the mRNA and protein expression of HIF-1α and CD133 before and after chemoradiotherapy. Spearman rank correlation was used to analyze the correlation between HIF-1α and CD133 mRNA expression. Univariate and logistic multivariate analyses were used to determine the factors related to pathological complete response (pCR). Logistic regression analysis and Cox's proportional hazard model were used to determine factors related to overall survival and recurrence-free survival.The expression of HIF-1α and CD133 mRNA was correlated with pT, ypTNM, pCR, recurrence and metastasis of rectal cancer, while not correlated with sex, age and BMI of patients. HIF-1α mRNA expression was positively correlated with CD133 mRNA expression (=0.579,=0.000). Immunohistochemistry analysis showed that residual cancer cells strongly expressing HIF-1α also expressed CD133 strongly. Univariate analysis showed that HIF-1α mRNA and CD133 mRNA were significantly correlated with pCR (=0.001,=0.022, respectively). Multivariate analysis showed that HIF-1α and CD133 mRNA expression were independent prognostic factors of pCR (=0.012,=0.047, respectively). Cox regression analysis showed that the expression of HIF-1α mRNA and CD133 mRNA were independent predictors of recurrence-free survival and overall survival (=0.025,=0.033, respectively).The study indicates that HIF-1α and CD133 can predict pathological complete remission and survival of patients with locally advanced rectal cancer.
AC133 Antigen ; analysis ; genetics ; Biomarkers, Tumor ; analysis ; Chemoradiotherapy ; Disease-Free Survival ; Female ; Humans ; Hypoxia-Inducible Factor 1, alpha Subunit ; analysis ; genetics ; Male ; Neoadjuvant Therapy ; Neoplasm Grading ; Neoplasm Metastasis ; diagnosis ; Neoplasm Recurrence, Local ; epidemiology ; genetics ; Neoplasm, Residual ; genetics ; Prognosis ; Proportional Hazards Models ; Rectal Neoplasms ; chemistry ; epidemiology ; genetics ; therapy ; Survival Rate

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

Cancer stem cells (CSCs) have tumor initiation, self-renewal, metastasis and chemo-resistance properties in various tumors including colorectal cancer. Targeting of CSCs may be essential to prevent relapse of tumors after chemotherapy. Phosphatidylinositol-3-kinase (PI3K) and mammalian target of rapamycin (mTOR) signals are central regulators of cell growth, proliferation, differentiation, and apoptosis. These pathways are related to colorectal tumorigenesis. This study focused on PI3K and mTOR pathways by inhibition which initiate differentiation of SW620 derived CSCs and investigated its effect on tumor progression. By using rapamycin, LY294002, and NVP-BEZ235, respectively, PI3K and mTOR signals were blocked independently or dually in colorectal CSCs. Colorectal CSCs gained their differentiation property and lost their stemness properties most significantly in dual-blocked CSCs. After treated with anti-cancer drug (paclitaxel) on the differentiated CSCs cell viability, self-renewal ability and differentiation status were analyzed. As a result dual-blocking group has most enhanced sensitivity for anti-cancer drug. Xenograft tumorigenesis assay by using immunodeficiency mice also shows that dual-inhibited group more effectively increased drug sensitivity and suppressed tumor growth compared to single-inhibited groups. Therefore it could have potent anti-cancer effects that dual-blocking of PI3K and mTOR induces differentiation and improves chemotherapeutic effects on SW620 human colorectal CSCs.
AC133 Antigen/genetics/metabolism ; Animals ; Antineoplastic Agents/pharmacology/therapeutic use ; Cell Differentiation/*drug effects ; Cell Line, Tumor ; Cell Survival/drug effects ; Chromones/pharmacology/therapeutic use ; Colorectal Neoplasms/drug therapy/metabolism/pathology ; Humans ; Imidazoles/pharmacology/therapeutic use ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Morpholines/pharmacology/therapeutic use ; Neoplastic Stem Cells/cytology/drug effects/metabolism ; Paclitaxel/pharmacology/therapeutic use ; Phosphatidylinositol 3-Kinases/*antagonists & inhibitors/metabolism ; Quinolines/pharmacology/therapeutic use ; SOXB1 Transcription Factors/genetics/metabolism ; Signal Transduction/*drug effects ; Sirolimus/pharmacology/therapeutic use ; TOR Serine-Threonine Kinases/*antagonists & inhibitors/metabolism ; Xenograft Model Antitumor Assays

OBJECTIVETo investigate the association of CD133 expression in rectal cancer tissues with neoadjuvant chemoradiotherapy (nCRT) and tumor regression grading (TRG) after nCRT.

METHODSRadical resected rectal cancer specimens and clinicopathological data of 105 patients, including 60 men and 45 women with median age of 59 years, diagnosed as locally advanced rectal cancer in Peking Union Medical College Hospital from January 2008 to December 2014 were collected retrospectively. Thirty-nine and 66 cases were histologically classified as good-moderate and poor differentiation respectively. Sixty-eight and 37 cases were clinically graded as stage I(-II( and III(-IIII( in preoperative assessment respectively. NCRT was administered in 61 cases before surgery (nCRT group). The nCRT consisted of preoperative pelvic radiotherapy using 50 Gy (2 Gy once, for 25 sessions) with FOLFOX regimen (5-fluorouracil plus oxaliplatin) for 2-3 cycles or XELOX regimen (capecitabine plus oxaliplatin) for 2 cycles. Patients underwent surgery after 6 courses of nCRT, and then received the same previous chemotherapy regimen. In nCRT group, biopsy specimens before nCRT were obtained in 45 cases. Forty-four cases received surgery alone without nCRT (surgery alone group). CD133 expression was tested by immunohistochemical Envision two-step methods. The histological TRG evaluation was performed in the nCRT group. TRG score 0-2 was defined as insensitivity to nCRT, whereas TRG score 3-4 was defined as sensitivity. CD133 expression in rectal cancer samples before and after nCRT was compared. Association of CD133 expression with TRG after nCRT was examined.

RESULTSNo significant differences of baseline parameters were found between nCRT group and surgery alone group (all P>0.05). The positive rate of CD133 in nCRT group was 70.4%(43/61,) which was significantly higher than that in surgery alone group (47.7%, 21/44)(χ(2)=5.566, P=0.018) and that in biopsy samples before nCRT group (44.4%, 20/45)(χ(2)=7.287, P=0.007). Twenty-two cases (36.1%, 22/61) in nCRT group had TRG score of 3-4 . Among these 22 cases, 11 cases were negative CD133, and constituted 61.1% (11/18) of all CD133-low expression cases in nCRT group, whereas the other 11 cases were positive CD133, and constituted 25.6%(11/43) of all CD133-high expression cases in nCRT group (χ(2)=6.974, P=0.008).

CONCLUSIONThe CD133 expression up-regulates markedly in rectal cancer after nCRT and nCRT may have potential positive modulation on CD133 expression. CD133-positive cancer reveals lower response to nCRT, suggesting CD133 may be a potential target for improving efficacy of nCRT in rectal cancer.

AC133 Antigen ; metabolism ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Chemoradiotherapy ; Deoxycytidine ; analogs & derivatives ; therapeutic use ; Female ; Fluorouracil ; analogs & derivatives ; therapeutic use ; Humans ; Leucovorin ; therapeutic use ; Male ; Middle Aged ; Neoadjuvant Therapy ; Neoplasm Staging ; Organoplatinum Compounds ; therapeutic use ; Rectal Neoplasms ; metabolism ; therapy

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

OBJECTIVE: To explore the effect of bone marrow mesenchymal stem cells (BM-MSCs) on angiogenesis in rats after brain injury. 
 METHODS: Brain injury model of rats was established with freely fall method. A total of 50 Sprague-Dawley (SD) rats were randomly divided into a transplanted group and a control group (n=25 in each group). BM-MSCs were injected in lateral ventricle in the transplanted group, and normal saline was injected in the control group. Modified method for neurological deficit scores (mNSS) was performed at the 1, 3, 7, 21, 14 d after the operation. Flow cytometry were performed to detect CD34 and CD133 double-labeled peripheral blood cells in preoperative or 3, 6, 12, 24 h, and 3, 7 d after the operation. Expression of neuron-specific enolase (NSE) and CD31 in the brain tissues near injury area was detected by immunohistochemical SP method. 
 RESULTS: There was significant difference in the MNSS scores between the 2 groups (F=5.997, P<0.05), and the difference at the different time points in each group was significant (F=37.106, P<0.01). The mNSS scores in the control group were higher than those in the transplanted group at the 7, 14, 21 d after the operation (P<0.05). The CD34 and CDl33 double positive cells (DPCs) were present in rats' peripheral blood. DPCs's numbers in peripheral blood in the control group were declined at 3 h after the sugery, they were increased and reached the highest point at 6 h after the surgery, and decreased gradually and reached normal levels at 24 h after the surgery. The same tendency was achieved in the transplanted group, and the DPCs's numbers were increased until 24 h after the surgery, which were significantly higher in the transplanted group than those in the control group at 24 h after the surgery (P<0.05). The NSE expression in the transplanted group was significantly greater than that in the control group in 7 and 14 d after the surgery (P<0.05). The expression of CD31 in the transplanted group was significantly higher than that in the control group in 3 and 7 d after the surgery (P<0.05).
 CONCLUSION BM-MSCs transplantation can increase the number of peripheral blood endothelial progenitor cells after traumatic brain injury in rats and sustain for 24 h, which in turn up-regulate the angiogenesis and neuronal marker, and improve the neurological function.
AC133 Antigen ; Animals ; Antigens, CD ; metabolism ; Antigens, CD34 ; metabolism ; Brain Injuries ; therapy ; Glycoproteins ; metabolism ; Hematopoietic Stem Cells ; cytology ; Mesenchymal Stem Cell Transplantation ; Neovascularization, Physiologic ; Peptides ; metabolism ; Phosphopyruvate Hydratase ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo explore expressions of CD133, E-cadherin and Snail in human epithelial ovarian cancer (EOC) and elucidate their relationship with the clinicopathologic features and prognosis of the patients.

METHODSThe expression of CD133, E-cadherin and Snail were detected by immunohistochemical staining in 150 specimens of EOC and 50 specimens of benign ovarian epithelial tumor tissues.

RESULTSThe positivity rates of CD133, E-cadherin and Snail protein in EOC were 58.7%, 60.7% and 32.7%, respectively, significantly different from the rates in benign epithelial tumor tissues (10%, 8.0%, and 70%, respectively; P<0.05). The expressions of CD133, E-cadherin and Snail in EOC were significantly correlated with abdominal organ and lymphnode metastases and FIGO stage (P<0.01). E-cadherin expression was inversely correlated with Snail and CD133 expression (r=-0.545 and -0.570, P<0.01), and the latter two were positively correlated (r=0.599, P<0.01). Overexpressions of CD133 and Snail and a decreased expression of E-cadherin were all related to a poor prognosis of the patients (P<0.05). FIGO stage and expressions of CD133, E-cadherin and Snail were all independent prognostic factors of EOC (P<0.05).

CONCLUSIONThe expressions of CD133, E-cadherin and Snail are related to lymph node metastasis, clinical stage, and prognosis of EOC. Combined detection of these indexes provides important evidence for predicting the progression and prognosis of EOC.

AC133 Antigen ; Antigens, CD ; metabolism ; Cadherins ; metabolism ; Disease Progression ; Female ; Glycoproteins ; metabolism ; Humans ; Lymphatic Metastasis ; Neoplasms, Glandular and Epithelial ; metabolism ; pathology ; Ovarian Neoplasms ; metabolism ; pathology ; Peptides ; metabolism ; Prognosis ; Snail Family Transcription Factors ; Transcription Factors ; metabolism

OBJECTIVETo investigate whether SIS3, a specific inhibitor of Smad3 phosphorylation, can reverse the stemness of multidrug-resistant(MDR) hepatocellular carcinoma cells.

METHODSMDR HCC Huh7.5.1/ADM cell lines were developed by exposing parental cells to stepwise increasing concentrations of ADM. CCK-8 assay was used to determine the cellular sensitivity of various anticancer drugs. Flow cytometry (FCM) was used to analyze the expression level of cancer stem cell marker CD133. Clone formation assay and mouse subcutaneous xenograft tumors were used to investigate the tumorigenicity in vitro and in vivo. Western blotting (WB) was used to analyze the changes of expressions of CD133, Smad3, Bcl-2, Bax and p-Smad3 in different conditions.

RESULTSADM treatment of HCC cells in vitro resulted in a development of subline, Huh7.5.1/ADM cells, with CSC phenotypes: stable MDR phenotype (besides ADMc Huh7.5.1/ADM cells were also more resistant to some other anticancer drugs including VCR, MMC and CTX ) (IC50: 0.215 ± 0.018 vs. 0.123 ± 0.004, 0.145 ± 0.009 vs. 0.014 ± 0.002, 1.021 ± 0.119 vs. 0.071 ± 0.006, 27.007 ± 1.606 vs. 1.919 ± 0.032) (unit: µg/ml) (P<0.05). Huh7.5.1/ADM cells enriched the cancer stem-like cell fraction (CD133-positive subpopulation) (76.06 ± 2.948% vs. 25.38 ± 4.349%) (P<0.05), had stronger tumorigenicity in vivo and colony formation ability, and activated the Smad3 activity. Inhibition of Smad3 activity by SIS3 decreased stemness of the Huh7.5.1/ADM cells: CD133-positive subpopulation (48.49 ± 2.304% vs. 76.06 ± 2.948%) (P<0.05); ADM IC50: (0.112 ± 0.019 vs. 0.215 ± 0.018), VCR IC50 (0.065 ± 0.013 vs. 0.145±0.009), MMC IC₅₀ (0.749 ± 0.121 vs. 1.021 ± 0.119), CTX IC50 (10.576 ± 1.248 vs. 27.007 ± 1.606) (unit: µg/ml) (P<0.05), and decreased tumorigenicity and colony formation ability.

CONCLUSIONSIS3 as a specific inhibitor of Smad3 signal is involved in the stemness of multidrug resistant hepatocellular carcinoma cells.

AC133 Antigen ; Animals ; Antibiotics, Antineoplastic ; pharmacology ; Antigens, CD ; metabolism ; Carcinoma, Hepatocellular ; drug therapy ; metabolism ; pathology ; Doxorubicin ; pharmacology ; Drug Resistance, Neoplasm ; Glycoproteins ; metabolism ; Heterografts ; Humans ; Isoquinolines ; pharmacology ; Liver Neoplasms ; drug therapy ; metabolism ; pathology ; Mice ; Neoplasm Proteins ; metabolism ; Neoplastic Stem Cells ; drug effects ; Peptides ; metabolism ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Pyridines ; pharmacology ; Pyrroles ; pharmacology ; Smad3 Protein ; antagonists & inhibitors ; metabolism ; Tumor Stem Cell Assay ; bcl-2-Associated X Protein ; metabolism

OBJECTIVETo investigate the presence of cancer stem cells (CSCs) exist in non-small cell lung cancer (NSCLC) and explore the relationship among the expressions of CD133, Notch1, and vascular endothelial growth factor (VEGF) and their relations with the clinicopathological parameters of the patients.

METHODSA total of 305 specimens of NSCLC and 80 normal lung tissue specimens were analyzed for CD133, Notch1, and VEGF protein expressions by immunohistochemical staining.

RESULTSIn NSCLC specimens, the positivity rates of CD133, Notch1, and VEGF were 48.9%, 43.9%, and 45.6%, respectively, significantly higher than those in normal lung tissues (10.0%, 15.0%, and 0%, respectively, P<0.01). The expression levels of CD133, Notch1, and VEGF proteins were significantly correlated with the tumor grades, lymph node metastasis, TNM stages, and postoperative survival time of the patients (P<0.01). A positive correlation was found among the expression levels of CD133, Notch1, and VEGF proteins. Kaplan-Meier survival analysis showed a significantly lower overall mean survival time of the patients positive for CD133, Notch1, and VEGF than that of the negative patients (P<0.001). Cox regression analysis suggested that positive expressions of CD133 and Notch1 were independent prognostic factors of NSCLC (P<0.05).

CONCLUSIONSCD133, Notch1, and VEGF may play important roles in the occurrence, progression, invasion, and metastasis of NSCLC. CD133 and Notch1 have important values for predicting the prognosis and evaluating disease progression of the patients.

AC133 Antigen ; Antigens, CD ; metabolism ; Carcinoma, Non-Small-Cell Lung ; metabolism ; Glycoproteins ; metabolism ; Humans ; Kaplan-Meier Estimate ; Lung ; metabolism ; Lung Neoplasms ; metabolism ; Lymphatic Metastasis ; Neoplastic Stem Cells ; metabolism ; Peptides ; metabolism ; Prognosis ; Receptor, Notch1 ; metabolism ; Regression Analysis ; Survival Rate ; Vascular Endothelial Growth Factor A ; metabolism

OBJECTIVETo investigate the expressions of WWOX and CD133 in colorectal cancer (CRC) and their relationship with the clinicopathologic characteristics of CRC.

METHODSThe expressions of WWOX and CD133 proteins were examined by immunohistochemistry in 174 specimens of CRC tissues and 80 normal colorectal mucosa tissues.

RESULTSThe positivity rates of WWOX and CD133 proteins were 41.4% and 53.4% in CRC tissues, respectively, significantly different from the rates in normal colorectal mucosa tissues (87.5% and 5.0%, respectively; P<0.05). WWOX and CD133 protein expressions were signi- ficantly correlated with the histological grades of the tumors, depth of invasion, lymph node metastasis, and Duke's stages (P<0.05). Spearman analysis showed a negative relationship between the WWOX expression and CD133 expression (P<0.05). Kaplan-Meier survival analysis showed that the overall survival time of CRC patients with a positive expression of WWOX was longer than that of patients with a negative expression of WWOX; the overall survival time of patients with a positive expression of CD133 was shorter than that of the negative patients (P<0.05). COX regression analysis identified positive expressions of WWOX and CD133 protein and Duke's stage as the independent prognostic factors of CRC.

CONCLUSIONAbnormal expressions of WWOX and CD133 might be involved in the initiation, development, invasion, and metastasis of CRC. A combined detection of WWOX and CD133 can help in predicting the progression and prognosis of CRC.

AC133 Antigen ; Antigens, CD ; metabolism ; Colorectal Neoplasms ; metabolism ; Disease Progression ; Glycoproteins ; metabolism ; Humans ; Immunohistochemistry ; Kaplan-Meier Estimate ; Lymphatic Metastasis ; Oxidoreductases ; metabolism ; Peptides ; metabolism ; Prognosis ; Tumor Suppressor Proteins ; metabolism ; WW Domain-Containing Oxidoreductase