1.Gene expression of transforming growth factor beta receptor II in the epidermis of pathological scar.
Ming-Rui CHEN ; Gang AN ; Shun-Li LIU ; Feng-Cai WEI
Chinese Journal of Burns 2012;28(4):278-281
OBJECTIVETo study the gene expression of transforming growth factor beta receptor II (TbetaR II) in pathological scar.
METHODSTwenty samples of pathological scar were collected from 20 burn or trauma patients hospitalized in the General Hospital of Ji'nan Military Command from 2007 to 2009. Twenty specimens of epidermal layer were obtained from the middle portion and the edge of pathological scars. Twenty normal skin specimens which were located more than 10 cm away from the lesion sites of 20 patients were collected as self-controls. Serum from 1-2 mL whole blood were obtained from each of the 20 patients for second self-control. Eight normal skin specimens from 8 patients without pathological scar, discarded from un-related operations, were also collected as negative-control. Positive expressions of TbetaR II in three different skin specimens were determined with biotin-streptavidin-peroxidase staining. Gene expressions of TbetaR II in all specimens were compared with PCR-single strand conformation polymorphism analysis and gene sequencing. Data were processed with Fisher's exact test.
RESULTSPositive expression of TbetaR II in pathological scar epidermis was lower than that in normal skin specimen of patients with pathological scar or normal skin specimen of patients without pathological scar, and TbetaR II was mainly located in the basal layer of epidermis. Positive expressions of TbetaR II were seldom found in acanthocytes, granular cells, and cuticle or even non-existing. No abnormality of TbetaR II was found in normal skin epidermis or serum samples of pathological scar patients or normal skin epidermis of patients without pathological scar. TbetaR II expressing in 8 specimens of epidermis of pathological scar showed abnormal electrophoresis pattern at poly A fragments hand and loss of one A base in DNA fragment (P = 0.044).
CONCLUSIONSThere may he abnormal gene expression of TbetaR II in pathological scar epidermis. Replantation of epidermis of scar may increase the risk of scar recurrence, while replantation of normal skin of patients with scar on wound may not increase the risk of scar recurrence.
Adolescent ; Adult ; Child ; Child, Preschool ; Cicatrix ; metabolism ; pathology ; Epidermis ; metabolism ; Female ; Gene Expression ; Humans ; Male ; Middle Aged ; Protein-Serine-Threonine Kinases ; genetics ; metabolism ; Receptors, Transforming Growth Factor beta ; genetics ; metabolism ; Young Adult
2.Negative Regulatory Effect of T-Lymphocytes on the Expansion of Human Bone Marrow Hematopoietic Cells In Vitro
Zhi-Gang TIAN ; Jian-Hua ZHANG ; Jun-Fu WANG ; Cai ZHANG ; Rui SUN
Journal of Experimental Hematology 2001;9(4):310-313
In this research, the regulatory effects of T-lymphocytes on the expansion of hematopoietic cells in human bone marrow were studied. Anti-CD3 McAb and anti-CD8 McAb were used to eliminate the T-lymphocytes in bone marrow MNCs. Cell surface antigens were analysed by flow cytometry. Hematopoietic cells expanded with hematopoietic growth factors (HGFs) in a liquid culture system and the number of CD34(+) cell, CFU-GM and CFU-GEMM were determined. After cultured with HGFs for 20 days the total number of cells expanded by 75.8, 79.6, 77.4 and 67.0 folds respectively in three experimental groups (anti-CD3 McAb, anti-CD8 McAb and anti-CD3 + anti-CD8 McAb) and control group (MNC of bone marrow). The number of CFU-GM in the four groups were 173.67 +/- 18.90, 165.33 +/- 26.58, 170.33 +/- 21.50 and 79.67 +/- 8.33 respectively. The number of CFU-GEMM in the four groups were 431.33 +/- 34.56, 370.33 +/- 42.10, 386.67 +/- 10.02 and 177.67 +/- 26.86 respectively. There were significant differences in the number of CFU-GM and CFU-GEMM between experimental groups and control group. The results showed that the T-lymphocytes in bone marrow could inhibit the expansion of hematopoietic cells in vitro and the formation of CFU-GM and CFU-GEMM. The regulatory mechanism was to be explored.
3.Effect of curcumin on radiosensitization of CNE-2 cells and its mechanism.
Qi-Rui WANG ; Hao-Ning FAN ; Zhi-Xin YIN ; Hong-Bing CAI ; Meng SHAO ; Jian-Xin DIAO ; Yuan-Liang LIU ; Xue-Gang SUN ; Li TONG ; Qin FAN
China Journal of Chinese Materia Medica 2014;39(3):507-510
OBJECTIVETo investigate the effect of curcumin (Cur) on radiosensitivity of nasopharyngeal carcinoma cell CNE-2 and its mechanism.
METHODThe effect of curcumin on radiosensitivity was determined by the clone formation assay. The cell survival curve was fitted by Graph prism 6. 0. The changes in cell cycle were analyzed by flow cytometry (FCM). The differential expression of long non-coding RNA was detected by gene chip technology. Part of differentially expressed genes was verified by Real-time PCR.
RESULTAfter 10 micro mol L-1 Cur had worked for 24 h, its sensitization enhancement ratio was 1. 03, indicating that low concentration of curcumin could increase the radiosensitivity of nasopharyngeal carcinoma cells; FCM displayed a significant increase of G2 phase cells and significant decrease of S phase cells in the Cur combined radiation group. In the Cur group, the GUCY2GP, H2BFXP, LINC00623 IncRNA were significantly up-regulated and ZRANB2-AS2 LOC100506835, FLJ36000 IncRNA were significantly down-regulated.
CONCLUSIONCur has radiosensitizing effect on human nasopharyngeal carcinoma CNE-2 cells. Its mechanism may be related to the changes in the cell cycle distribution and the expression of long non-coding IncRNA.
Cell Cycle ; drug effects ; radiation effects ; Cell Line, Tumor ; Cell Survival ; drug effects ; radiation effects ; Curcumin ; pharmacology ; Gene Expression Regulation, Neoplastic ; drug effects ; radiation effects ; Humans ; RNA, Long Noncoding ; genetics ; Radiation Tolerance ; drug effects
4.Effects of anastrozole on lipid metabolism in Chinese postmenopausal women with breast cancer.
Han-hong LU ; Qing LI ; Bing-he XU ; Pin ZHANG ; Peng YUAN ; Jia-yu WANG ; Rui-gang CAI
Chinese Journal of Oncology 2011;33(7):520-525
OBJECTIVEThe aim of this study was to evaluate the effect of anastrozole, a new generation aromatase inhibitor, on the lipid metabolism in postmenopausal Chinese women with early breast cancer, and observe the adverse reactions as well.
METHODSPostmenopausal women with early breast cancer patients took anastrozole 1 mg per day. The lipid profiles of total cholesterol, triglyceride, low density lipoprotein, and high density lipoprotein were assessed before taking the drug, 3 months, 6 months after taking medication, and later once a year, until the end of medication or follow-up. Patients taking lipid-lowering drugs were excluded. The adverse reactions during the process of taking medication was followed-up by telephone.
RESULTSTwo hundred and eighty-five postmenopausal breast cancer patients took part in the trial from Jan. 2003 to Jun. 2009. All patients had completed primary surgery and demonstrated a postmenopausal status. ER or PR positivity was confirmed by histopathology. Taking the medication from a minimum of one year to a maximum of 5 years, with a median time of 3.61 years. During the medication time, anastrozole significantly increased the levels of low density lipoprotein-cholesterol after 6 months of treatment, continuing to 5 years, from (3.08 ± 0.90) mmol/L to (3.59 ± 0.59) mmol/L, with a maximal increase of 18.2% higher than that before medication. Anastrozole significantly increased the levels of total cholesterol and high density lipoprotein-cholesterol after 1 years of treatment. Anastrozole significantly reduced the levels of triglycerides after 1 years of treatment. Anastrozole showed no significant effect on serum lipids in the patients with pre-existing hyperlipidemia. A more significant effect on blood lipids was observed in patients aged ≥ 60-years than that in patients less than 60 years of age. The rate of other adverse events were similar to that reported in foreign patients.
CONCLUSIONSFor the postmenopausal patients with breast cancer, taking anastrozole may lead to an abnormal lipid metabolism. Anastrozole significantly increases the levels of low density lipoprotein-cholesterol, total cholesterol and high density lipoprotein-cholesterol, and significantly reduces the level of triglycerides. The rate of other adverse events were similar to that reported in foreign patients. it is suggested that the blood lipid levels should be regularly assessed in patients with long-term anastrozole treatment. The rate of other adverse events similar to that reported with foreign patients, and patients tolerate this treatment well.
Age Factors ; Aged ; Aged, 80 and over ; Antineoplastic Agents, Hormonal ; therapeutic use ; Aromatase Inhibitors ; therapeutic use ; Breast Neoplasms ; blood ; complications ; drug therapy ; surgery ; Chemotherapy, Adjuvant ; Cholesterol ; blood ; Cholesterol, HDL ; blood ; Cholesterol, LDL ; blood ; Female ; Follow-Up Studies ; Humans ; Hyperlipidemias ; blood ; complications ; Lipid Metabolism ; drug effects ; Lipids ; blood ; Middle Aged ; Neoplasm Staging ; Nitriles ; therapeutic use ; Postmenopause ; Triazoles ; therapeutic use ; Triglycerides ; blood
5.Effect of RNA interference inhibition to expression of CD133 on tumor cell biological characteristics in KATO-III CD133(+) cells of human gastric cancer.
Shou-lian WANG ; Ji-wei YU ; Cheng CAI ; Rui-qi LU ; Ju-gang WU ; Xiao-chun NI ; Bo-jian JIANG
Chinese Journal of Gastrointestinal Surgery 2013;16(9):889-894
OBJECTIVETo investigate the changes in proliferation, invasiveness, clone sphere formation and chemosensitivity of human gastric cancer cell lines of KATO-III CD133(+) cells transfected with small interfering RNA (siRNA) against CD133 gene.
METHODSCD133(+) cells of KATO-III cell lines were isolated by magnetic activated cell sorting (MACS). CD133 siRNA was designed and synthesized, and then transfected into KATO-III CD133(+) cells. Cell fluorescence counting under confocal laser scanning microscope was used to determine the transfection efficiency after transfection with the CD133 FITC-siRNA. The knock-down effect of the CD133 gene and expression of epithelial-mesenchymal transition (EMT)-related factors were detected by RT-PCR and Western blotting. Cell counting kit-8 assay (CCK-8), transwell chamber and colony sphere forming assay were performed to measure the variation of cell proliferative, invasive, colony formation viability and chemosensitivity to 5-FU after the above-mentioned treatment.
RESULTSThe transfection efficiency was (87.7±8.1)%. The CD133 mRNA and protein expression levels in the interference group were lower than those in negative control group. Twenty-four, 48 and 72 hours after transfection, cells proliferation activity was significantly inhibited in the interference group compared with negative control group, (all P<0.01). Seventy-two hours after transfection, compared with negative control group, cells proliferation activity was reduced by (52.1±8.0)%. The invasive cell number reduced (41.7±6.0 vs. 130.3±11.0, P<0.05) and clone formation rate decreased significantly [(24.3±4.3)% vs. (45.1±6.4)%, P<0.01] in the interference group. EMT-related gene E-cadherin protein expression increased, while the Snail and N-cadherin protein expression reduced in the interference group (all P<0.01). The cells sensitivity to 5-FU was significantly enhanced in the interference group, and the cell inhibition rate of 5-Fu was (62.4±3.3)%, higher than that in negative control group [(21.5±2.2)%, P<0.01].
CONCLUSIONSThe expression of CD133 gene plays an important role in cell proliferation, invasiveness, colony formation and resistance to chemotherapy of KATO-III CD133(+) gastric cancer cells. It suggests that CD133 can be used as one of surface markers for detection of gastric cancer stem cells. Inhibition of CD133 expression may be a promising way for gastric cancer biotherapy.
AC133 Antigen ; Antigens, CD ; genetics ; metabolism ; Cell Line, Tumor ; Cell Movement ; Cell Proliferation ; Fluorouracil ; pharmacology ; Glycoproteins ; genetics ; metabolism ; Humans ; Peptides ; genetics ; metabolism ; RNA Interference ; RNA, Small Interfering ; genetics ; Stomach Neoplasms ; genetics ; metabolism ; pathology ; Transfection
6.Endobronchial metastases from breast cancer: a clinicopathological and survival analysis.
Jian LI ; Bing-he XU ; Jia-yu WANG ; Qing LI ; Pin ZHANG ; Peng YUAN ; Fei MA ; Ying FAN ; Rui-gang CAI ; Qiao LI
Chinese Journal of Oncology 2012;34(5):394-397
OBJECTIVEEndobronchial metastases (EBM) secondary to extrapulmonary solid malignant tumors are rare but may occur. The most common extrathoracic malignancies associated with EBM are colorectal, renal and breast cancer. This study aimed to evaluate the clinicopathological aspects of EBM from breast cancer and the prognosis of the patients.
METHODSClinicopathological data of 11 cases diagnosed as EBM from breast cancer treated in our hospital from 2003 to 2010 were re-evaluated. Their symptoms, recurrence interval, radiological features, histopathological properties, and prognosis were assessed.
RESULTSEleven cases were diagnosed by bronchoscopic bronchial biopsy. The median interval from diagnosis of breast cancer was 57 months (range: 11 - 189 mo). All patients had other proven metastases when the EBM was diagnosed. The most frequently observed symptoms were cough (8 cases). Interestingly, two patients were asymptomatic. Hilar mass (5 cases) was a common radiological finding. No disaccordance between the hormone receptor status in the primary and metastatic lesions in these patients was found. The median survival after EBM diagnosis was 21 months (range: 6 - 36) with four patients still alive and one of these four patients was surviving more than 7 years.
CONCLUSIONSOn average, EBM is diagnosed about 5 years after the diagnosis of breast cancer, which is a relatively long lead time, but the median survival time is short, as 21 months in our group. The treatment plan must be individualized, because in some cases, long-term survival can be expected.
Adult ; Antineoplastic Agents ; therapeutic use ; Breast Neoplasms ; drug therapy ; pathology ; radiotherapy ; surgery ; Bronchial Neoplasms ; drug therapy ; secondary ; Carcinoma, Ductal, Breast ; drug therapy ; pathology ; radiotherapy ; secondary ; surgery ; Chemotherapy, Adjuvant ; Female ; Follow-Up Studies ; Humans ; Lymphatic Metastasis ; Mastectomy, Modified Radical ; Middle Aged ; Nitriles ; therapeutic use ; Radiotherapy, Adjuvant ; Retrospective Studies ; Survival Analysis ; Triazoles ; therapeutic use
7.Unfavorable pathological complete response rate of neoadjuvant chemotherapy epirubicin plus taxanes for locally advanced triple-negative breast cancer.
Yi YIN ; Pin ZHANG ; Bing-he XU ; Bai-lin ZHANG ; Qing LI ; Peng YUAN ; Rui-Gang CAI ; Jia-yu WANG ; Xiang WANG ; Xiao-zhou XU
Journal of Huazhong University of Science and Technology (Medical Sciences) 2013;33(2):262-265
Anthracycline-Taxane chemotherapy is widely used in neoadjuvant treatment for breast cancers. However, there is limited data reported in patients with triple negative breast cancer (TNBC). Here, we evaluated the pathologic responses and survival of neoadjuvant epirubicin and taxanes chemotherapy in patients with locally advanced TNBC to provide some useful information for clinical practice. A total of 43 patients with locally advanced TNBC were enrolled in this study. Patients were administered with epirubicin 75 mg/m(2) plus paclitaxel 175 mg/m(2) or docetaxel 75 mg/m(2) every 3 weeks for at least 2 cycles. The primary endpoint was pathologic complete response (pCR), which was defined as no residual invasive cancer, or only carcinoma in situ in both the excised breast and axillary lymph node, while relapse-free survival (RFS) and overall survival (OS) were secondary endpoints. Thirty-nine (90.7%) patients were at clinical stages IIB-IIIC. Thirty-seven (86%) completed 4-6 cycles of preoperative chemotherapy, and objective response rate (ORR) was 81.4% (35/43). Forty-two patients underwent radical surgery subsequently. The pCR rate was 14.3% (6/42). The most common adverse events in neoadjuvant chemotherapy were nausea/vomiting (88.4%, 38/43) and neutropenia (88.4%). After a median follow-up period of 34.0 months, 3-year RFS and OS rate was 53.6% and 80.1%, respectively. All events of recurrence and death occurred in non-pCR patients, in whom the 3-year RFS and OS rates were 44.3% and 76.6%, respectively. This study suggest that neoadjuvant chemotherapy with epirubicin plus taxanes has a relatively low pCR rate and high early recurrence risk in locally advanced TNBC, which indicates the necessity for more efficacious treatment. Further study is needed to validate these results.
Adult
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Aged
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Antineoplastic Combined Chemotherapy Protocols
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administration & dosage
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Chemoradiotherapy, Adjuvant
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methods
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Disease-Free Survival
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Epirubicin
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administration & dosage
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Female
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Humans
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Male
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Middle Aged
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Neoadjuvant Therapy
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methods
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Neoplasm Recurrence, Local
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pathology
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prevention & control
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Neoplasm, Residual
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pathology
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prevention & control
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Paclitaxel
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administration & dosage
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Taxoids
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administration & dosage
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Treatment Failure
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Treatment Outcome
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Triple Negative Breast Neoplasms
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drug therapy
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pathology
8.The observation of blood serum creatinine and phosphorus during long-term adefovir dipivoxil treatment in chronic hepatitis.
Jing-Hua TIAN ; Yan-Qun HE ; Xiao-Yan MA ; Rui-Hong LI ; Shu-Feng ZHANG ; Gang WAN ; Hao-Dong CAI
Chinese Journal of Hepatology 2013;21(3):239-240
Adenine
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analogs & derivatives
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therapeutic use
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Adolescent
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Adult
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Aged
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Creatinine
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blood
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Female
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Hepatitis B, Chronic
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blood
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drug therapy
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Humans
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Male
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Middle Aged
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Organophosphonates
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therapeutic use
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Phosphorus
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blood
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Retrospective Studies
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Young Adult
9.Sorting of CD133(+) subset cells in human gastric cancer and the identification of their tumor initiating cell-like properties.
Rui-qi LU ; Ju-gang WU ; Guo-cai ZHOU ; Hai-guang JIANG ; Ji-wei YU ; Bo-jian JIANG
Chinese Journal of Gastrointestinal Surgery 2012;15(2):174-179
OBJECTIVETo sort CD133(+) subset cells in human gastric cancer (GC) and to identify their tumor initiating cell-like properties.
METHODSThe tissues of GC and normal tissues adjacent to GC were obtained from 50 patients. Samples were stained for CD133 by immunohistochemistry. Likewise, assessments of CD133 were undertaken by Western blot. Flow cytometry was used to determine the proportion of CD133(+) cells in four GC cell lines therein the KATO-III was sorted by magnetic activated cell sorting (MACS) method. The growing characteristics and the tumorigenic ability of CD133(+) cells were evaluated in vitro and in vivo. Meanwhile, the growth of single cells in suspension culture was observed and expression of stem cell-specific marker were determined using reverse transcription-polymerase chain reaction (RT-PCR).
RESULTSThe expression of CD133 was demonstrated on the cell membranes in the mucosa and submucosa of primary GC, which were higher than those in the normal gastric tissues adjacent to cancer (P<0.05). Four GC cell lines including KATO-III, SGC-7901, AGS and MKN-45 were found to contain (28 ± 2)%, (17 ± 2)%, (6 ± 2)%, and (4 ± 2)% of CD133(+) cells respectively. In addition, the purity of CD133(+) cells isolated from KATO-III by MACS was (91 ± 3)% and up to(95 ± 2)% after 1-week culture. CCK-8 detection showed that population doubling time of the CD133(+) cells was (21 ± 3)h, significantly shorter than that of the CD133(-) cells[(40 ± 8)h, P<0.05]. Notably, there was a remarkable difference of tumor formation rate between CD133(+) cells (100%), non-sorted cells (80%), and CD133(-) cells(0). The average mass and volume of tumor in group of CD133(+) cells was larger and heavier than those in non-sorted cells (P<0.05, P<0.05). Furthermore, the single cell proliferated well, formed the big sphere and semi-quantitative RT-PCR showed expression of stem cell markers such as Oct-4, Nanog, Sox-2, Musashi-1 and EGFR.
CONCLUSIONSCD133 protein expression in primary lesions is higher than those in the normal gastric tissues. CD133(+) subset cells can be isolated, purified, and amplified in human GC, and possess some properties including the ability of self-renewal, proliferation, and higher tumorigenic ability in vivo and can express some stem cell markers.
AC133 Antigen ; Adult ; Aged ; Aged, 80 and over ; Animals ; Antigens, CD ; metabolism ; Cell Line, Tumor ; Female ; Glycoproteins ; metabolism ; Humans ; Male ; Mice ; Mice, Inbred BALB C ; Middle Aged ; Neoplastic Stem Cells ; metabolism ; Peptides ; metabolism ; Stomach Neoplasms ; metabolism ; pathology ; Xenograft Model Antitumor Assays
10.Down regulation of multidrug resistance-associated protein 4 expression by RNA interference enhances radiosensitivity of colorectal carcinoma cell lines in vitro.
Zhi-qi YU ; Chang ZHANG ; Rui CAI ; Xin-yuan LAO ; Hao WANG ; Xian-hua GAO ; Yi-fang HAN ; Xiao-qing ZHANG ; Guang-wen CAO ; Chuan-gang FU
Chinese Journal of Gastrointestinal Surgery 2012;15(1):67-71
OBJECTIVETo investigate the effect of multidrug resistance-associated protein 4 (MRP4) expression on the radiosensitivity of colorectal carcinoma cell lines in vitro.
METHODSThe vector of shRNA for RNA interference was constructed and then transfected into HCT116 cell line to steadily down-regulate the expression of MRP4. HCT116 cells were divided into 3 groups including the CON group(non-transfected), NC group (negative control virus was added), and KD group (RNAi target was added for transfection). To test the effectiveness of RNA interference, real-time polymerase chain reaction and Western blot were used to measure the expression pattern of MRP4 at both mRNA and protein levels, respectively. For the examination of the effect of RNA interference of MRP4 on the radiosensitivity, flow cytometry was used to calculate the rate of apoptotic cells 24 h after 4 Gy radiation. Proliferation of the cells was measured via MTT assay at different time points.
RESULTSShRNA plasmid was successfully constructed. Transfection of this constructed vector into HCT116 cell line caused steady silencing of MRP4 expression (HCT116-KD). MRP4 mRNA and protein expression were significantly down-regulated following RNA interference(P<0.05). Twenty-four hours after radiation, the apoptosis rate of KD cell line was (71.7±0.8)%, significantly higher than that in the CON group [(56.1±0.9)%] and NC group[(59.8±0.8)%](P<0.05). Fourty-eight hours and 72 hours after radiation, the proliferation was significantly inhibited in KD cells compared to the control groups(P<0.05).
CONCLUSIONSExpression of MRP4 is closely related to radio-tolerance of colorectal carcinoma. Down-regulation of MRP4 expression by RNA interference enhances radiosensitivity of colorectal carcinoma cell lines in vitro. MRP4 may be an effective molecular marker for predicting the radiosensitivity of colorectal carcinoma.
Colorectal Neoplasms ; genetics ; metabolism ; Down-Regulation ; HCT116 Cells ; Humans ; Multidrug Resistance-Associated Proteins ; genetics ; RNA Interference ; Radiation Tolerance ; genetics