1.Study on the chemoresistance of CD44+/CD24+ Siha cells to cisplatin and its mechanisms
Hong LIU ; Yujing WANG ; Lei BIAN ; Haili LI ; Zhaohui FANG ; Xiaohua WU ; Jianxin CHENG
China Oncology 2015;(10):785-790
Background and purpose:One of the reasons why cancer cells are resistant to chemotherapy is the existence of cancer stem cells. The purpose of this study was to investigate the chemoresistance of CD44+/CD24+ Siha cells to cisplatin and its mechanisms.Methods:Siha cells were cultivatedin vitro. The CD44+/CD24+ Siha cells were sorted out by fluorescence activated cell sorter (FACS) andin vitro proliferation was detected by MTT assay after treatment with the different concentrations of cisplatin. The cell apoptosis rate was detected by flow cytometry after 10 μg/mL cisplatin acted on CD44+/CD24+ Siha cells for 24, 48 and 72 h. The relative mRNA and protein expressions of Bcl-2, Oct-4 and ABCG2 were detected by quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot, respectively. Results:The survival rates of CD44+/CD24+ Siha cells treated with different concentrations of cisplatin (0.1, 1, 5, 10, 15 and 20 μg/mL) were higher than those of their parental Siha cells [(88.42±1.51)%vs (92.87±1.5)%, (79.94±1.05)%vs (84.72±1.09)%, (69.78±0.81)%vs (75.13±2.86)%, (58.97±0.70)%vs (65.79±2.71)%, (49.60±0.88)%vs (52.10±0.52)%, (45.13±0.69)%vs (48.84±1.02)%,P<0.05]. Compared with their parental Siha cells, the apoptosis rates of CD44+/CD24+ Siha cells were lower after 10 μg/mL of cisplatin acting on them for 24, 48 and 72 h, respectively [(3.05±0.16)%vs (5.17±0.27)%, (17.94±2.02)%vs (32.60±4.28)% and (40.14±3.01)%vs (56.62±5.32)%,P<0.05]. The results from both qRT-PCR and Western blot indicated that Oct-4, ABCG2 and Bcl-2 were highly expressed on CD44+/CD24+ Siha cells. A significant difference was found in Oct-4, ABCG2 and Bcl-2 expression between CD44+/CD24+Siha cells and their parental cells (P=0.015<0.05).Conclusion:CD44+/CD24+ Siha cells could be resistant to apoptosis induced by cisplatin and expressed high levels of cancer stem cell markers such as Oct-4 and ABCG2. This study lays the basis for useful isolation and further targeted therapy of cervical cancer stem cells.
2.Resistance of CD44+/CD24+ expressing cervical cancer cells on apoptosis induced by X-ray irradiation
Hong LIU ; Yujing WANG ; Lei BIAN ; Qianying ZHANG ; Zhaohui FANG ; Xiaohua WU ; Jianxin CHENG
Chinese Journal of Radiological Medicine and Protection 2015;35(9):647-651
Objective To explore whether CD44 +/CD24 + expressing cervical cancer cells are resistant to X-ray irradiation and investigate the underlying mechanism.Methods Cervical cancer cell line (Siha) was cultured in vitro and the CD44 +/CD24 + expressing cells were sorted with a flow cytometer.The cells were irradiated with 8,16 and 30 Gy of 6 MV X-rays.Colony formation test was used to evaluate the radiosensitivity of CD44 +/CD24 + expressing cervical cancer cells.Cell morphology was observed by electronmicroscopy,cell apoptosis was analyzed with a flow cytometer and also verified with a DNA ladder assay.Gene expression was determined by RT-PCR.Results After radiation,the ratio of CD44 +/CD24 + cells significantly increased.Compared to Siha cells,the radiosensitivity of CD44 +/ CD24 + cells decreased (t =93.99-400.45,P <0.05),and the expressions of bcl-2,survivin and Oct4 mRNA increased in CD44 +/CD24 + cells (t =221.35,941.65,82.27,P <0.01).Both apoptotic body and specific DNA ladder pattern were observed in cells but not in the CD44 +/CD24 + Sihacells which had no obvious morphological changes of apoptosis.Conclusions The CD44 +/CD24 + expressing cervical cancer cells are resistant to X-rays due to expression of anti-apoptosis factors.
3.Effects on cellular immunity caused by intravenous infusion of allogenic rhesus mesenchymal stem cells
Chuanbo FAN ; Zhaohui WANG ; Lei WANG ; Kaixun HU ; Lihui LIU ; Qiyun SUN ; Li BIAN ; Qingchao WU
Journal of Leukemia & Lymphoma 2011;20(9):550-553
ObjectiveTo study the changes of cellular immunity caused by intravenous infusion of allogenic rhesus mesenchymal stem cells (MSCs).MethodsMSCs were isolated and cultured.Then the immunomodulatory effects after MSCs infusion were evaluated by means of peripheral blood counts,mixed lymphocyte reaction (MLR) and analysis of lymphocytic subgroup. ResultsMSCs of rehsus were successfully cultivated. No acute toxicities or GVHD were observed in recipients. No obvious changes of peripheral blood counts were present. Recipients A2, A3, A4 were administered with MSC by 4.0 ×105/kg, 1.0 ×106/kg, 2.0×106/kg respectively and relative reaction (RR) of MLR decreased 14 days post MSCs infusion: from 46±2.6 %to 40.4±1.73 % (F =10.19, P =0.023), from (40.9±2.3) % to (33±2.1) % (F =2.593, P =0.013), from 48.3±2.0 % to 39±1.0 % (F =28.431, P =0.003) respectively. The decrease degree (ARR) was positively related to the amount of MSCs(F =27.413, P =0.038). RR was restored within 30 days post MSCs infusion. After MSCs infusion, CD3+ CD3+CD4+ and CD3+CD8+ T-lymphocytes decreased in recipient A4, who was administered with the largest number of MSCs, and restored within 30 days. ConclusionMSCs infusion without any other treatment could temporarily inhibit immunity of T lymphocytes in MLR and the immunity inhibition was positively related to the amount of MSCs.The specific immunological characteristics of MSCs were demonstrated with extensive prospect in clinical research.
4.Transferring the radialis proper digital nerve of index finger and its dorsal branch to repair the thumb nerve evulsion
Qichao LI ; Zhangmin LIU ; Zhaohui BIAN ; Jianzhong JIANG ; Wenchai SHAO ; Xiaobo FENG
Chinese Journal of Primary Medicine and Pharmacy 2017;24(24):3694-3696
Objective To investigate the clinical curative effect of the surgical method of transferring the radialis proper digital nerve of damaged index finger and its dorsal branch to repair the thumb nerve evulsion.Methods 13 patients with thumb nerve evulsion were treated.There were 8 males and 5 females with an average age of 28 years (ranged 18-52 years old).The injuries were caused by machine twist (8 cases),gear(4 cases),electric saw (1 case).And thumb rotational avulsion amputation (10 cases),thumb incompleteness amputation(3 cases).The time from injury to admission was 1-3.5 h (mean 2.2 h).The average time from injury to admission was (1.1 ± 1.5) h.The amputate level of skin was at the juxtra-articular of metacarpophalangeal joint.The amputate level of bone was at the base of proximal phalanx (6 patients) and metacarpophalangeal joint (4 patients),interphalangeal joint(3 patients).Using transferring the radialis proper digital nerve of index finger without reimplantation and its dorsal branch to.repair the both side injuries of thumb nerve evulsion.According to routine method to repair digital bone,veins,arteries and tendons.Results All 13 chases were followed up for 6 months to 2 years and 7 months postoperatively,with an average of 22 months.The wounds and incisions at donor sites were healed by first intention.All 13 cases of thumb reimplantation were successful.Two-point discrimination of ulnaris finger pulp was 2 to 6mm,average 4.2mm,and the radialis was 5 to 9mm,average 7.8mm.Sensory function was rated as S4(the ulnaris in 12 cases and the radialis in 2 cases) and S3 + (the radialis in 11 cases and the ulnaris in 1 cases).Conclusion Transferring the radialis proper digital nerve of damaged index finger and its dorsal branch to the digital nerve on the neighboring thumb is a simple and effective method to restore sensory function of the pulp.
5.MiR-139-5p inhibits migration and invasion of colorectal cancer by downregulating AMFR and NOTCH1.
Mingxu SONG ; Yuan YIN ; Jiwei ZHANG ; Binbin ZHANG ; Zehua BIAN ; Chao QUAN ; Leyuan ZHOU ; Yaling HU ; Qifeng WANG ; Shujuan NI ; Bojian FEI ; Weili WANG ; Xiang DU ; Dong HUA ; Zhaohui HUANG
Protein & Cell 2014;5(11):851-861
MicroRNAs (miRNAs) that exert function by posttranscriptional suppression have recently brought insight in our understanding of the role of non-protein-coding RNAs in carcinogenesis and metastasis. In this study, we described the function and molecular mechanism of miR-139-5p in colorectal cancer (CRC) and its potential clinical application in CRC. We found that miR-139-5p was significantly downregulated in 73.8% CRC samples compared with adjacent noncancerous tissues (NCTs), and decreased miR-139-5p was associated with poor prognosis. Functional analyses demonstrated that ectopic expression of miR-139-5p suppressed CRC cell migration and invasion in vitro and metastasis in vivo. Mechanistic investigations revealed that miR-139-5p suppress CRC cell invasion and metastasis by targeting AMFR and NOTCH1. Knockdown of the two genes phenocopied the inhibitory effect of miR-139-5p on CRC metastasis. Furthermore, the protein levels of the two genes were upregulated in CRC samples compared with NCTs, and inversely correlated with the miR-139-5p expression. Increased NOTCH1 protein expression was correlated with poor prognosis of CRC patients. Together, our data indicate that miR-139-5p is a potential tumor suppressor and prognostic factor for CRC, and targeting miR-139-5p may repress the metastasis of CRC and improve survival.
Animals
;
Base Sequence
;
Cell Line, Tumor
;
Cell Movement
;
genetics
;
Colorectal Neoplasms
;
genetics
;
pathology
;
therapy
;
Down-Regulation
;
Female
;
Gene Expression Profiling
;
Gene Expression Regulation, Neoplastic
;
HCT116 Cells
;
HEK293 Cells
;
Humans
;
Male
;
Mice, Inbred BALB C
;
Mice, Nude
;
MicroRNAs
;
genetics
;
Middle Aged
;
Neoplasm Invasiveness
;
RNA Interference
;
Receptor, Notch1
;
genetics
;
metabolism
;
Receptors, Autocrine Motility Factor
;
genetics
;
metabolism
;
Reverse Transcriptase Polymerase Chain Reaction
;
Sequence Homology, Nucleic Acid
;
Survival Analysis
;
Xenograft Model Antitumor Assays
6.MicroRNA-638 inhibits cell proliferation by targeting phospholipase D1 in human gastric carcinoma.
Jiwei ZHANG ; Zehua BIAN ; Jialiang ZHOU ; Mingxu SONG ; Zhihui LIU ; Yuyang FENG ; Li ZHE ; Binbin ZHANG ; Yuan YIN ; Zhaohui HUANG
Protein & Cell 2015;6(9):680-688
MicroRNAs (miRNAs) are a type of small non-coding RNAs that are often play important roles in carcinogenesis, but the carcinogenic mechanism of miRNAs is still unclear. This study will investigate the function and the mechanism of miR-638 in carcinoma (GC). The expression of miR-638 in GC and the DNA copy number of miR-638 were detected by real-time PCR. The effect of miR-638 on cell proliferation was measured by counting kit-8 assay. Different assays, including bioinformatics algorithms (TargetScan and miRanda), luciferase report assay and Western blotting, were used to identify the target gene of miR-638 in GC. The expression of miR-638 target gene in clinical CRC tissues was also validated by immunohistochemical assay. From this research, we found that miR-638 was downregulated in GC tissues compared with corresponding noncancerous tissues (NCTs), and the DNA copy number of miR-638 was lower in GC than NCTs, which may induce the corresponding downregulation of miR-638 in GC. Ectopic expression of miR-638 inhibited GC cell growth in vitro. Subsequently, we identified that PLD1 is the target gene of miR-638 in GC, and silencing PLD1 expression phenocopied the inhibitory effect of miR-638 on GC cell proliferation. Furthermore, we observed that PLD1 was overexpressed in GC tissues, and high expression of PLD1 in GC predicted poor overall survival. In summary, we revealed that miR-638 functions as a tumor suppressor in GC through inhibiting PLD1.
3' Untranslated Regions
;
genetics
;
Apoptosis
;
genetics
;
Base Sequence
;
Cell Line, Tumor
;
Cell Proliferation
;
genetics
;
Down-Regulation
;
genetics
;
Humans
;
MicroRNAs
;
genetics
;
Phospholipase D
;
genetics
;
Prognosis
;
Stomach Neoplasms
;
diagnosis
;
enzymology
;
genetics
;
pathology
7.Correction to: MiR-139-5p inhibits migration and invasion of colorectal cancer by downregulating AMFR and NOTCH1.
Mingxu SONG ; Yuan YIN ; Jiwei ZHANG ; Binbin ZHANG ; Zehua BIAN ; Chao QUAN ; Leyuan ZHOU ; Yaling HU ; Qifeng WANG ; Shujuan NI ; Bojian FEI ; Weili WANG ; Xiang DU ; Dong HUA ; Zhaohui HUANG
Protein & Cell 2021;12(8):668-670