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Objective:To investigate the effects and its mechanism of long non-coding RNA (LncRNA) small nucleolar RNA host gene 6 (SNHG6) on proliferation, apoptosis and chemosensitivity of prostate cancer stem cells.Methods:CD44 + CD24 - tumor stem cells and non-CD44 + CD24 - cells were selected from prostate cancer cell PC-3 by flow separation technology, and the expression level of SNHG6 and microRNA (miR) -26a were detected by real-time fluorescence quantitative PCR. Cell proliferation was detected by 5-bromodeoxyuridine (Br-dU) , the apoptosis was detected by flow cytometry, the chemosensitivity of cells to cisplatin was detected by methyl thiazolyl tetrazolium (MTT) , and Western blot was used to detect the protein expressions of proliferating cell nuclear antigen (Ki-67) , B-cell lymphoma-2 (Bcl-2) and Bcl-2 associated X protein (Bax) ; moreover, the target relationships of SNHG6, miR-26a and zeste enhancer of zeste homolog 2 (EZH2) were detected by double luciferase reporter gene assay, and Western blot was used to detect the effect of miR-26a analog on EZH2 protein expression. Results:Compared with non-CD44 + CD24 - cells, the expression level of SNHG6 in CD44 + CD24 - cells was significantly higher ( P<0.05) ; compared with NC-siRNA group [ (1.00±0.06) %, (96.85±6.48) %, (0.72±0.06) %, (0.43±0.03) %, (5.32±0.15) %, (0.35±0.03) %], SNHG6 expression level (0.25±0.03) , cell proliferation activity [ (75.36±5.1) %], Ki67 (0.38±0.03) and Bcl-2 protein (0.21±0.02) expression levels in SNHG6-siRNA group were significantly lower, while miR-26a expression level, apoptosis rate [ (13.83±2.36) %] and Bax protein (0.48±0.03) expression level were significantly higher, and the sensitivity of the cells to cisplatin was significantly higher ( P<0.05) ; in addition, miR-26a was the target gene of SNHG6, EZH2 was the target gene of miR-26a, and miR-26a analog could reduce the expression level of EZH2 protein. Conclusions:SNHG6 is up-regulated in prostate cancer stem cells. Interfering SNHG6 expression can inhibit the proliferation of cancer stem cells, promote apoptosis, and enhance the sensitivity of cancer stem cells to cisplatin. The mechanism may be related to the targeting regulation of miR-26a/EZH2 axis.
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Protein phosphorylation is one of the main ways to activate protein bioactivity and make it participate in cell life activities. Researches have shown that approximately 30% of proteins in the human body are modified by phosphorylation at different levels and at different sites. If the protein phosphorylation modification level or site is abnormal, it will cause the occurrence and development of malignant tumors. Malignant tumors have always been a kind of diseases that endanger human life and health. According to statistics, as many as 18 million malignant tumors and more than 9.6 million deaths occur every year worldwide. With the continuous recognition on the abnormality of protein phosphorylation modification in tumorigenesis and development, the research and development of drugs for abnormality of protein phosphorylation modification has become a focus in the field of tumor therapy at present. Each traditional Chinese medicine(TCM) can be regarded as a natural molecular library, and it participates in the regulation of protein phosphorylation modification level with the advantages of multiple components and multiple targets, with slight side effect and low drug resistance, so TCM is one of the main sources of drug development for regulating protein phosphorylation modification levels. Through the search of multiple databases at home and abroad, it was found that certain monomers, parts extracted from TCM, single-TCM and TCM compounds can affect the tumor progression by regulating the level of protein phosphorylation and exert better anti-tumor effect. Based on the current research status of protein phosphorylation regulation by TCM at home and abroad, we found that TCM can inhibit tumor cell proliferation, invasion and metastasis, angiogenesis, and maintenance of stem cell stemness by regulating protein phosphorylation levels, and exert antitumor effects by promoting apoptosis. In order to clarify the molecular mechanism of TCM in regulating protein phosphorylation level and exerting antitumor effect, and provide evidences for the development and clinical application of antitumor TCM, the authors reviewed the mechanism of TCM in regulating protein phosphorylation level and exerting their antitumor effect from different ways in this paper.
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Lung cancer stem cells play an important role in chemoresistance and invasion and metastasis of lung cancer. Increasing evidence sustains that microRNAs(miRNAs)play a major role in regulating tumor angiogenesis,drug resistance and metastasis. Cancer stem cells are considered to be one of the reasons leading to cancer recurrence,metasta-sis and resistance to chemotherapy,therefore better understanding how miRNAs regulate gene expression in lung cancer stem cells will help identify cancer biomarkers and new therapeutic targets,and will help to develop better treatment strate-gies for lung cancer.
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Tumor cells have unique energy metabolism phenomena, namely high glucose absorption, aerobic glycolysis and high lactic acid production, which are characterized by down-regulation of related proteins involved in oxidative metabolism in tumor cells, and up-regulation of glucose transporters and monocarboxylate transporters. Studies have shown that drugs that target tumor cell glucose metabolism have the ability to selectively kill tumor cells, bringing new hope for tumor treatment. Tumor stem cells are considered to be the root cause of tumor recurrence, metastasis and poor prognosis, and their energy metabolism characteristics have not yet been agreed. Studies have shown that reversing the energy metabolism of tumor stem cells can increase their chemosensitivity. This article reviews recent studies on tumor and tumor stem cell glucose metabolism and the opportunities and challenges of tumor treatment through targeting glucose metabolism, which might provide new ideas and opportunities for clinical tumor therapy.
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Humans , Energy Metabolism , Glucose , Metabolism , Glycolysis , Lactic Acid , Metabolism , Neoplasms , Metabolism , Neoplastic Stem Cells , MetabolismABSTRACT
Objective: To test the application of flow cytometry technique to detect the redox status with genetically encoded fluorescent probe roGFP2, to compare it with laser scanning confocal microscope (LSCM), and to demonstrate the diversity of cellular redox status in HeLa and Panc-1 cell populations. Methods: Time lapse imaging with LSCM was performed in single cell transfected with roGFP2 probe to detect the dynamic changes of 405 nm/488 nm ratio (405/488 ratio). Flowcytometry technique was also performed in HeLa cells transfected with roGFP2 probe to detect the dynamic alterations of 405/488 ratio. The global cell population was analyzed and the subpopulations of different redox status were dissected. Flowcytometry technique was further applied in Panc-1 cells with different CD24 or CD44 marker to detect the dynamic alterations of 405/488 ratio of roGFP2 and identify the different redox status. Results: Time lapse imaging with LSCM showed that 405/488 ratio of roGFP2 dramatically changed in response to H2O2 in dose and time dependent manner at a single cell level. When dozens of cells were chosen, the average ratio showed increased and dynamic trend. Compared to LSCM, flow cytometry could detect the average of 405/488 ratio of roGFP2 as well. Meanwhile, with the application of flow cytometry the cell population can be divided into three subpopulations based on 405/488 ratios, most in oxidized and medium redox status, few in reduced status. Using flow cytometry, CD24+/CD44+ Panc-1 cells, pancreatic cancer stem cells, can be found to have overall lower 405/488 ratio and more percentage of subpopulation of reduced status under resting and stress condition. Conclusion: Flow cytometry technique can be applied to detect roGFP2 and has advantage in application to show the overall as well as diverse redox status in cell population. Flow cytometry detection of cellular redox status with genetically encoded probe can be a useful tool in tumor cell biology and developmental biology research.
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Objective·To test the application of flow cytometry technique to detect the redox status with genetically encoded fluorescent probe roGFP2,to compare it with laser scanning confocal microscope (LSCM),and to demonstrate the diversity of cellular redox status in HeLa and Pane-1 cell populations.Methods·Time lapse imaging with LSCM was performed in single cell transfected with roGFP2 probe to detect the dynamic changes of 405 nm/488 nm ratio (405/488 ratio).Flowcytometry technique was also performed in HeLa cells transfected with roGFP2 probe to detect the dynamic alterations of 405/488 ratio.The global cell population was analyzed and the subpopulations of different redox status were dissected.Flowcytometry technique was further applied in Panc-1 cells with different CD24 or CD44 marker to detect the dynamic alterations of 405/488 ratio of roGFP2 and identify the different redox status.Results·Thne lapse imaging with LSCM showed that 405/488 ratio of roGFP2 dramatically changed in response to H2O2in dose and time dependent manner at a single cell level.When dozens of cells were chosen,the average ratio showed increased and dynamic trend.Compared to LSCM,flow cytometry could detect the average of 405/488 ratio of roGFP2 as well.Meanwhile,with the application of flow cytometry the cell population can be divided into three subpopulations based on 405/488 ratios,most in oxidized and medium redox status,few in reduced status.Using flow cytometry,CD24+/CD44+ Panc-1 ceils,pancreatic cancer stem ceils,can be found to have overall lower 405/488 ratio and more percentage of subpopulation of reduced status under resting and stress condition.Conclusion·Flow cytometry technique can be applied to detect roGFP2 and has advantage in application to show the overall as well as diverse redox status in cell population.Flow cytometry detection of cellular redox status with genetically encoded probe can be a useful tool in tumor cell biology and developmental biology research.
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Tumor stem cells (TSCs), because of its close related to occurrence, development, metastasis, recurrence and drug resistance of tumor, are considered by research commnity as a root cell bank of the tumor. In other words, The key way to cure tumor may be to eradicate TSCs. The traditional surgery, chemotherapy and radiothera-py not only have few effect in killing TSCs, but result in recurrence and metastasis easily. However, TSCs can definitely killed by the immunotherapy targeting TSCs, for instance, targeting the surface antigens of TSCs', T cells tar-getting the TSCs, differentiation therapy and so on.
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Objective:To investigate the effect of Schisandra chinensis polysaccharide(SCP)on the growth of brain tumor stem cells(BTSCs),and to clarify the mechanism of inhibiting the growth of BTSCs of SCP. Methods:The primary human glioma cells were cultured,then the BTSCs were isolated by CD133 immunomagnetic sorting.The neural stem cell surface markers CD133 and Nestin were detected by immunofluorescence assay.The proliferation rate of BTSCs was examined by MTT assay.Annexin V-PI analysis was used to analyze the apoptotic rate of BTSCs.The expression levels of Bax,Bcl-2 and Caspase-3 proteins in BTSCs in various groups were detected by ELISA assay.Results:The results of immunofluorescence staining showed that the expressions of CD133 and Nestin were positive in BTSCs.Compared with control group,the proliferation rates of BTSCs in 200,400 and 800 mg·L-1SCP groups were decreased,especially in 400 and 800 mg·L-1SCP groups(P<0.05).The results of Annexin V-PI analysis showed that the apoptotic rate of BTSCs in 800 mg·L-1SCP group was increased compared with control group(P<0.05).The ELISA results showed that the expression levels of Bax in 200,400 and 800 mg·L-1SCP groups were significantly increased(P<0.05),and the values of Bax/Bcl-2 were significantly increased(P<0.05);compared with control group,the Bcl-2 expression level in the BTSCs in 800 mg·L-1SCP group was decreased(P<0.05).The expression level of Caspase-3 protein in 800 mg·L-1SCP group was also significantly increased compared with control group(P<0.01).Conclusion:SCP could inhibit the growth of BTSCs,and the induction of apoptosis may be one of mechanisms.
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To compare the therapeutic effects of different treatment methods on the nude mice bearing colon cancer HT29 cells. BalB/C nude mice colon cancer stem cell models were established and randomly divided into the following four groups, with 8 nude mice in each group: blank control group, DC-CIK group, Huaier group, and Huaier combined with DC-CIK group (combined treatment group). The mice in DC-CIK group and combined treatment group received 1×10⁶ DC-CIK cells treatment by tail vein injectionafter the tumor stem cells were inoculated for 4 days,2 times a week for three weeks. The mice in Huaier group and combined treatment group received intragastric administration at the dose of 20 g/60 kg body weight, 0.2 mL/time, once a day for a total of three weeks. The mice in control group received equal volume of normal saline. Tumor size and body weight of nude mice were measured every 2 days during treatment for three weeks in each group. After the treatment, the nude mice were sacrificed to measure the tumor weight and the tumor inhibition rate was calculated. The RT-PCR method was used to detect the expression levels of the key genes in the signal pathway. After the end of the treatment, the quality of the tumor in the Huaier group, DC-CIK group and combined treatment group was significantly lower than that in the control group; the quality in combined treatment group was significantly lower than that in Huaier group and DC-CIK group.Among them, the tumor inhibition rate reached 46.77% in the combined treatment group. In respect of changes in expression levels of key genes in the signaling pathway, the mRNA expression levels of key genes PI3KR1 and Akt in PI3K/Akt pathway, key genes Wnt1 and CTTNB1 in Wnt/-catenin pathway, and key genes Notch1, Notch2, Notch3 in Notch pathway in the combined treatment group were lower than those in DC-CIK group and Huaier group. The Huaier combined with DC-CIK group showed best therapeutic effect among different treatment methods for HT29 stemcell colon tumors in nude mice, providing a new idea for clinical treatment of colon cancer.
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Animals , Humans , Mice , Cell Line, Tumor , Colonic Neoplasms , Drug Therapy , Complex Mixtures , Pharmacology , Mice, Inbred BALB C , Mice, Nude , Neoplastic Stem Cells , Signal TransductionABSTRACT
Objective: To investigate the effect of Schisandra chinensis polysaccharide (SCP) on the growth of brain tumor stem cells (BTSCs), and to clarify the mechanism of inhibiting the growth of BTSCs of SCP. Methods: The primary human glioma cells were cultured, then the BTSCs were isolated by CD133 immunomagnetic sorting. The neural stem cell surface markers CD133 and Nestin were detected by immunofluorescence assay. The proliferation rate of BTSCs was examined by MTT assay. Annexin V-PI analysis was used to analyze the apoptotic rate of BTSCs. The expression levels of Bax, Bcl-2 and Caspase-3 proteins in BTSCs in various groups were detected by ELISA assay. Results: The results of immunofluorescence staining showed that the expressions of CD133 and Nestin were positive in BTSCs. Compared with control group, the proliferation rates of BTSCs in 200, 400 and 800 mg · L-1 SCP groups were decreased, especially in 400 and 800 mg · L-1 SCP groups (P<0.05). The results of Annexin V-PI analysis showed that the apoptotic rate of BTSCs in 800 mg · L-1 SCP group was increased compared with control group (P<0.05). The ELISA results showed that the expression levels of Bax in 200, 400 and 800 mg · L-1 SCP groups were significantly increased (P<0.05), and the values of Bax/Bcl-2 were significantly increased (P<0.05); compared with control group, the Bcl-2 expression level in the BTSCs in 800 mg · L-1 SCP group was decreased (P<0.05). The expression level of Caspase-3 protein in 800 mg · L-1 SCP group was also significantly increased compared with control group (P<0.01). Conclusion: SCP could inhibit the growth of BTSCs, and the induction of apoptosis may be one of mechanisms.
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Purpose To observe the expression of HIF-1αand HIF-2α in tumor stem cells and tumor tissues of small cell lung cancer (SCLC) and to explore their clinical significance.Methods The defined serum-free culture was used to enrich the third passage tumor spheres cells from H446 as the tumor stem cells.Real-time PCR was performed to determine the mR-NA expression level of HIF-1α and HIF-2α in H446 tumor stem cells.Immunofluorescence staining was performed to determine the protein expression level of HIF-1α and HIF-2α in H446 tumor stem cells.Immunohistochemistry SP method was used to detect the expression of HIF-1α and HIF-2αt in SCLC tissues.Results The mRNA expression level of HIF-2α was up-regulated in tumor stem cells.However,the mRNA expression level of HIF-1 α was down-regulated in tumor stem cells (P ＜ 0.05).The expression of HIF-2α protein was positive in tumor stem cells.In contrast,HIF-1α protein was negative in tumor stem cells.In SCLC tissues,the positive rate of HIF-1α was 46.7％ (28/60),and the positive rate of HIF-2α was 25％ (15/60).Correlation analysis showed that HIF-2α was positively correlated with SCLC stem cell marker uPAR,and they co-localized around necrotic regions.The expression of HIF-2α was closely related to tumor diameter and distant metastasis.In contrast,the expression of HIF-1α had no relationship with age,sexy,tumor size,lymph metastasis,pleural invasion and distant metastasis (P ＞ 0.05).Conclusion HIF-2α is up-regulated in SCLC stem cells and positively correlated with SCLC stem cell marker uPAR,which are associated with the tumor diameter and distant metastasis of SCLC patients,suggesting that the expression of HIF-2α may be related to SCLC stem-like characteristics.
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OBJECTIVE:To study the effect of Elemene emulsion combined with ganciclovir on invasive ability and TIMP-1 mRNA expression in C6 brain tumor stem cells. METHODS:Cells with 3 generations were isolated and cultured from rats'C6 ma-lignant glioma cell lines,and immunocytochemistry was adopted to detect the protein expressions of stem cell markers CD133,Nes-tin. 20 rats were randomly divided into blank control group(normal saline),ganciclovir group(intragastrically administrated,216 mg/kg),Elemene emulsion injection group (intravenously injected in tail,36 mg/kg) and combination group (intragastrically ad-ministrated 216 mg/kg of ganciclovir+intravenously injected 36 mg/kg of Elemene emulsion injection in tail),5 in each group, once every day,for 10 d. After 2 h of last administration,the blood of rats in each group was collected,serum was isolated and co-cultured 24 h with C6 BTSCs in in vitro culture medium. Boyden invasion test was used to detect the invasive ability of C6 BTSCs,and real-time quantitative polymerase chain reaction(RT-PCR)method was used to determine the TIMP-1 mRNA expres-sion of C6 BTSCs. RESULTS:Cells with 3 generations had obvious protein expressions in CD133 and Nestin,indicating they were BTSCs. Compared with blank control group,the cell invasion rates of C6 BTSCs in ganciclovir group,Elemene emulsion in-jection group and combination group were obviously decreased,TIMP-1 mRNA expression was obviously enhanced,with statisti-cal significances(P<0.05). Compared with ganciclovir group and Elemene emulsion injection group,the cell invasion rate of C6 BTSCs in combination group was obviously decreased,TIMP-1 mRNA expression was obviously enhanced,with statistical signifi-cances(P<0.05). CONCLUSIONS:Elemene emulsion injection combined with ganciclovir can obviously inhibit the cell invasion of C6 BTSCs,the mechanism may be associated with promoting the TIMP-1 generation,and combination use shows better effect than single use.
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Objective@#To explore the method of extracting chaperone antigen peptide complexes from gastric cancer stem cells and its immune function.@*Methods@#Gastric cancer stem cells and gastric cancer cells were screened by low temperature ultrasonic lysis. After salting out and dialysis, the lysate supernatant was processed with SDS-PAGE to analyze the expression of chaperone antigen peptide complexes, and then was separated and purified with CNBr-activated SepharoseTM 4B. Reverse high pressure liquid chromatography (HPLC), SDS-PAGE and Western blotting were used to analyze the purity and nature of the acquired albumen. Lymphocyte proliferation assay and lymphocytotoxicity assay were used to ditermine the immunological activity of the chaperone-antigen peptide complexes.@*Results@#The chaperone antigen peptide complexes of gastric cancer stem cells were prepared and identified successfully, of which the main components were the antigen peptides of HSP60, HSP70, HSP90 and HSP110. 0.75 μg and 1.00 μg HSP70-antigen peptide and 1.00 μg HSP90-antigen peptide activated lymphocytes significantly. Their A490 values were 0.26±0.03, 0.45±0.05 and 0.32±0.04, respectively, while the corresponding doses of HSP60-antigen peptide and HSP110-antigen peptide did not activate lymphocytes. The killing rates of 1.00 μg HSP70-antigen peptide and 1.00 μg HSP70 were (45.0±2.0)% and (16.0±2.0)%, respectively, showing a significant difference (P=0.012). Similarly, the killing rates of 1.00 μg HSP90-antigen peptide and 1.00 μg HSP90 were (36.0±5.0)% and (13.0±4.0)%, respectively, also showing a significant difference (P=0.048).@*Conclusions@#The amount of chaperone antigen peptide complexes in gastric cancer cells is extremely low, but it is obviously increased in gastric cancer stem cells. After purification, the chaperone antigen peptide complexes with high purity can be prepared. The extracted chaperone antigen peptide complexes have stronger immunogenicity, and can be used to make tumor vaccine in vitro, which may have a good application value in the targeted therapy of gastric cancer.
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OBJECTIVE: To detect the effect of TRAIL on the growth of brain tumor stem cells, study the role of Caspase-8 and Bcl-2 of TRAIL resistance. METHODS: Brain tumor stem cells were isolated by CD133 magnetic sorting, and the positive rates of CD133+ cells were detected by flow cytometry, the self-renewing capicity of brain tumor stem cells was examined by subneurosphere formation assay, and the percentage of cell death were examined by MTS assay, the expression of DR5, FADD, Caspase-8 and Bcl-2 proteins was detected by Western blot. RESULTS: The positive rates of CD133+ cells were 71% after CD133 magnetic sorting. Brain tumor stem cells grow as neurosheres and have the abilities to reform neuroshperes, the capacity of neurosphere formation was significant increased after TRAIL treatment or not. Brain tumor stem cells were dead after TRAIL treatments, Caspase-8 and Caspases inhibitor can block the cell death that induced by TRAIL (P < 0.05). The expression of Caspase-8 protein was decreased and Bcl-2 protein was increased in brain tumor stem cells (P < 0.05). CONCLUSION: Brain tumor stem cells may response to TRAIL resistance, the reason is the lower expression of Caspase-8 protein and over expression of Bcl-2 protein which is unable to activate Caspase-8.
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Objective: To explore the inhibitory mechanism of polypeptide extract from scorpion venom (PESV) on sarcoma S180. Methods: Thirty mice were implanted with S180 cells and randomly divided into three groups: control group, PESV group, and rapamycin (RAPA) group with 10 mice in each group. Then the tumor volume growth curve was drawn and the tumor inhibitory rate (IR) was calculated. The morphological changes of the tumor tissue were observed by HE staining. The protein expression levels of Beclin1, MAP1LC3A, and CD133 were detected using immunohistochemical assay. Western blotting was applied to detecting the expression of Beclin1, MAP1LC3A, and CD133 in tumor tissue of mice in each group. Results: The growth of sarcoma S180 transplanted tumor was inhibited more obviously in the PESV group and RAPA group than that in the control group. The IR in the PESV and RAPA groups were 17.9% and 25.0%, respectively (P 180, the mechanisms might be associated with promoting the expression of autophagic relative factors, Beclin1 and MAP1LC3A as well as inhibiting the expression of CD133.
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Objective To enrich breast cancer stem cells of breast cancer cell lines MCF-7 and MDA-MB-231 through culturing mammospheres,and to detect the expression differences of gene of breast cancer stem cells makers.Methods The MCF-7 and MDA-MB-231 cell lines were cultured by serum and serum-free medium.The proportion of CD44+/CD24-and CD133+ cancer stem cells was measured in cells derived from mammosphere cells or monolayer culture cells by flow cytometry,and the expression of CD44,CD24,CD133,ALDH3A1,ABCG2 and CXCR4 mRNA were detected by RT-PCR.Results Flow cytometry analysis indicated that the CD44+/CD24-low proportion of the MCF-7 mammosphere cells was higher than its adherent culture cells (P ＜ 0.05),and CD133+ proportion had no difference between them (P ＞ 0.05).However,the CD44+/CD24-/low proportion of the MDA-MB-231 mammosphere cells was lower than its adherent culture cells (P ＜ 0.05),while CD133+ proportion was significantly higher than its adherent cultured cells (P ＜ 0.05).RT-PCR analysis suggested that the expression of CD44 and ABCG2 increased obviously in MCF-7 microspheres (P＜ 0.05),and the expression of CD24,CD133,ALDH3A1 and CXCR4 had no significant difference between the mammosphere cells and adherent culture cells (P ＞ 0.05).The CD24,CD133,ABCG2,ALDH3A1 and CXCR4 increased obviously in MDA-MB-231 microspheres.On the contrary,the CD44 decreased (P ＜ 0.05).The expression of CD44 and CD24,CD133 and ALDH3A1 had significant differences in the microspheres of MCF-7 and MDA-MB-231 cells (P ＜ 0.05).Conclusion The related cancer stem cells gene expression is different in the microspheres of human breast cancer cell line,which indicates that the different subtypes of breast cancer may be derived from different origins.
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Objective To observe inhibitory effects of Ginsenoside on the proliferation cultured stem cells of human rectal carcinoma in vitro.Methods Tumor stem cells of rectal carcinoma in human were cultured in vitro and divided into saline water group,reltitrexed group(3mg/mL),and Ginsenoside(25μg/mL)plus reltitrexed(3mg/mL) group,with dosage of qd and for 2 weeks.Cells growth was observed under the microscope,and immunofluorescence staining was used to detect CD +133 expression of rectal carcinoma cells.MTT colorimetric method was taken to measure inhibitory of Ginsenoside on proliferation of tumor stem cells of rectal carcinoma.Apoptosis of tumor stem cells of rec-tal carcinoma lead by Ginsenoside was observed by DAPI staining.Results Part of cells were growing up like ball and CD +133 immunofluorescence staining was positive.Inhibitory role of Ginsenoside plus reltitrexed on CD +133 cells of rectal carcinoma appeared on the first day,which was bigger than that of retitrexed by MTT method (P =0.03).Com-pared with the retitrexed group,Gisenoside combined with reltitrexed reduced CD +133 cells expression of rectal carcino-ma,in which optical density and area density of CD +133 cells were obviously decreased with statistical significance (P =0.007,P =0.006,respectively).Conclusion Gisenoside plays the remarkably inhibitory role on proliferation of CD +133 cells of rectal carcinoma.
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Objective To identify the differential expressed genes of human glioma stem cells (GSCs) and human neural stem cells (NSCs) by gene chip technology. Methods Human HOA5.1 OneArray microarray (including 29 186 genes) was adopted and hybridized with probes which were prepared from the total RNAs of GSCs and NSCs. Differential expressed genes between the GSCs and NSCs were assayed after scanning oligonucleotide microarray with ScanArray 4000, and some of these genes such as DCX, PTGS2, SCGN, GAD2, OTX2, PEG 10 and NRXN3 were verified by real-time-Q-PCR method. Results Compared with the genes in normal NSCs, 1372 down-regulated and 1501 up-regulated genes in GSCs were revealed by means of microarray, and these genes were associated with axon guidance, cell cycle, cell adhesion, immune-inflammatory responses and cancer-related signal pathways. The results of qRT-PCR were consistent with that of microarray. Conclusions Multiple genes play important roles in development of glioma. This study may provide new clues for the targeted therapy of malignant glioma.
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Objective To evaluate the effect of hepatocellular carcinoma suppressor gene-1 (HCCS1) on the cancer stemlike phenotype, and invasive and metastatic ability of liver tumor cells. Methods Cells of human liver tumor cell line MHCC-97H were divided into three groups: normal control group, HCCS1 group and Ad-LacZ group. Cells in normal control group received no specific treatment, and cells in HCCS1 group and Ad-LacZ group were infected with Ad-HCCS1 and Ad-LacZ respectively, with infestation rate of 100:1. Real-time PCR and Western blotting were used to assess the mRNA and protein expression of HCCS1. FACS was used to identify the proportion of CD133 and CD90 positive cells. ELISA was employed to measure the levels of VEGF in the supernatant. Western blotting was used to detect the E-cadherin and fibronectin expression. Transwell assay was conducted to test the invasive and metastatic ability for the tumor cells. Results The mRNA and protein expression of HCCS1 in the HCCS1 group were higher than those in normal control group and Ad-LacZ group (P<0.01). Compared with normal control and Ad- LacZ group, the proportion of CD133 and CD90 positive cells, the content of VEGF in the supernatant, and the number of cells infiltrating Transwell membrane significantly decreased, while the expression of E-cadherin and fibronectin significantly increased in the HCCS1 group (P<0.01). Conclusion HCCS1 could inhibit cancer stem-like phenotype, and invasive and metastatic ability of MHCC-97H cells.
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Photoacoustic flow cytometry (PAFC) is a novel flow cytometry which integrates high-pulse repetition-rate lasers,fast signal acquisition algorithms and focused ultrasound transducers to assess deep vessels.The technical principle is that the cells in blood or lymph flow are irradiated with several laser beams with different wavelengths,then laser-induced PT effects are detected by corresponding schematics.PAFC is characterized by its high efficiency,no invasion and real-time detection,which makes it possible to detect tumor cells in circulation or in lymphatic system dynamically and in real time.So for,it is considered as one of the most promising techniques in cancer research.This article will address the principle,application and several problems of PAFC.