OBJECTIVETo explore and identify the non-coding RNAs related to tumors.

METHODSWe used RT-PCR and Northern blot to analyze non-coding RNAs in tumor tissues and cell lines.

RESULTSTwo predicted non-coding RNAs were confirmed to be expressed in cancer tissues and cell lines by RT-PCR and DNA sequencing. We detected the expression of two non-coding RNA transcripts by Northern blot. The length of NC28 was about 1800 nt, and that of NC119 was about 1200nt.

CONCLUSIONSNC28 and NC119 have a tumor-associated expression pattern. The non-coding RNAs may play a role in the development of tumors.

Cell Line, Tumor ; Humans ; Neoplasms ; metabolism ; RNA, Untranslated ; biosynthesis

Carbonic anhydrase IX (CAIX) is a transmembrane protein that is specifically overexpressed on the surface of hypoxic tumor cells. With the function of regulating the acidity of tumor cells both inside and outside, CAIX is closely related to tumor proliferation, invasion and metastasis. Therefore, CAIX is a promising target for tumor imaging and therapy. Herein, we summarized recent advances in CAIX-based tumor imaging, therapy and theranostics, and prospected future applications of using CAIX as an anti-tumor target.
Carbonic Anhydrase IX ; Carbonic Anhydrases/metabolism* ; Cell Line, Tumor

OBJECTIVETo detect miR-143 expression in human nasopharyngeal carcinoma (NPC) cell lines and explore the role of miR-143 in regulating the adhesion ability of NPC cells.

METHODSFluorescence quantitative RT-PCR was used to detect miR-143 expression levels in 5 NPC cell lines (CEN1, CNE2, HONE1, 5-8F, and 6-10B) and an immortalized human nasopharyngeal epithelial cell line (NP69). The retrovirus plasmid pMSCV-puro-miR-143 was constructed and the packaged retroviruses pMSCV-puro and pMSCV-puro-miR-143 were infected in 5-8F cells to establish a cell line with stable miR-143 overexpression, whose adhesion ability was tested with adhesion assay.

RESULTSThe expression of miR-143 was down- regulated in the NPC cell lines, and the highly metastatic NPC cell line 5-8F had a expression of only 3.0% of the control level, as compared with the level of 63.59% in the tumorigenic but nonmetastatic NPC cell line 6-10B. The transfected 5-8F cells showed a 1080-fold increase of miR-143 expression (P<0.05) with a significantly lowered adhesion ability.

CONCLUSIONmiR-143 plays a role in regulating the invasiveness and metastasis of NPC, and overexpression of miR-143 causes a significant reduction of the adhesion ability of the highly metastatic NPC cell line 5-8F.

Carcinoma ; Cell Adhesion ; Cell Line, Tumor ; Cell Movement ; Humans ; MicroRNAs ; metabolism ; Nasopharyngeal Neoplasms ; metabolism ; pathology

On-line analysis and control are critical for the optimization of product yields in animal cell culture. The close monitor of viable cell number helps to gain a better insight into the metabolism and to refine culture strategy. In this study, we use the oxygen uptake rate (OUR) to estimate the number of viable cell and the OUR-based feed-back control strategy for nutrients feeding to improve the efficiency of cell culture. A hybridoma cell line (HAb18) was cultured in fed-batch and perfusion model using serum free medium in 5L CelliGen Plus bioreactor (NBS Co., American) and 5L Biostat B bioreactor (Braun Co., Germany). The system and the method for online monitoring OUR in bioreactors, based on the dynamic measurement of dissolved oxygen (DO), were developed. The method of on-line cell concentration estimation was established based on the relationship between the growth of the hybridoma and the uptake rate of oxygen. This method was then used to determine OUR and the concentrations of cell, antibody, glucose, lactate, glutamine and ammonia in the bioreactors at given times. The relationship between OUR and nutrients metabolism was studied and OUR-based feed-back control strategy, which used the state deltaOUR = 0 as the regulation point, was established and used to control the rates of nutrients or medium feeding rate in the perfusion culture. The results showed that there was close relationship between OUR, concentration of live cells, productivity of antibody and consumption of glutamine. The sudden decrease in OUR may be caused by glutamine depletion, and with different delay times, the viable cell concentration and antibody productivity also decreased. The further analysis revealed the linear relationship between OUR and the density of live cells in the exponential growth phase as qOUR = (0.103 +/- 0.028) x 10(-12) mol/cell/h. These findings can be applied to the on-line detection of live cell density. Our study also indicated that by adjusting the perfusion rate with OUR-based feed-back control strategy, it is feasible to continuously increase in viable cell density and antibody concentration in the perfusion culture.
Bioreactors ; Cell Culture Techniques ; methods ; Cell Line, Tumor ; Cell Proliferation ; Humans ; Hybridomas ; metabolism ; Oxygen ; metabolism

OBJECTIVETo investigate the expression of miR-9 in B lymphocytes, B cell lymphoma and classical Hodgkin's lymphoma (cHL) cell lines and its significance.

METHODSCD19(+) B lymphocytes were sorted from normal lymph node by magnetic beads. Total cellular micro-RNA was extracted from cHL cell line L428, B cell lymphoma cell lines Ly1 and Ly10 (diffuse large B cell lymphoma), Raji cells (Burkitt's lymphoma) and CD19(+) B lymphocytes, respectively. These micro-RNAs were separately transformed into cDNA by reverse transcription. The expression levels of miR-9 were measured by fluorescence quantitative PCR. In situ hybridization was used to detect the expression of miR-9 in cell lines.

RESULTSThe expression of miR-9 was high in L428 cells (104.44 ± 1.61), and low in cell lines of B cell lymphoma (Ly1: 2.17 ± 0.38; Ly10: 1 ± 0.015; Raji: 2.65 ± 0.89), and extremely low in CD19(+) B lymphocytes (0.0026 ± 0.00040). Compared with that in the other cell lines, the expression of miR-9 in L428 cells was statistically significant (P < 0.05). miR-9 localized in the cytoplasm diffusely and strongly in L428, but scattered and slightly with some prominent distribution around the nuclear membranes in Ly1 and Ly10, and only weakly in Raji.

CONCLUSIONSmiR-9 highly expressed in cHL cell line and might be a molecular marker for diagnosis and treatment of cHL.

B-Lymphocytes ; metabolism ; Cell Line, Tumor ; Cell Lineage ; Hodgkin Disease ; metabolism ; pathology ; Humans ; Lymphoma, B-Cell ; metabolism ; pathology ; MicroRNAs ; metabolism

The study was aimed to investigate the expression of Notch signal molecules in human lymphoma cells and its significance. Raji, Maver, Z138 and Jurkat cell lines were used in the study. RT-PCR was used to determine the expression of Notch signaling molecules in these lymphoma cells. Flow cytometry was used to detect the apoptosis and cell-cycle of the lymphoma cells induced by different concentrations of gamma secretase inhibitor DAPT. CCK-8 was used to detect the proliferation of the lymphoma cells treated by DAPT. The results showed that the expression of Notch molecules in the four cell lines was different. Notch1 and Notch2 were found to be expressed in the four lymphoma cell lines, Notch3 predominantly expressed in Jurkat cells, Notch4 only expressed in Raji cells weakly and Hes1 only expressed in Raji and Jurkat cells. Treatment with DAPT could increase the apoptosis ratio of Raji and Jurkat cells. Besides, DAPT could significantly inhibit the proliferation of Raji and Jurkat cells by inducing the cell cycle arrest in G(1) phase, but the effect of DAPT on Maver and Z138 cells was not obvious. The activity of Notch pathway could be inhibited by DAPT treatment through down-regulating the expression of Notch target gene Hes1. It is concluded that the abnormal expression and activation of Notch signal pathway play an important role in the proliferation of lymphoma cells. Notch may be likely a new target for the therapy of lymphoma.
Apoptosis ; Cell Cycle ; Cell Line, Tumor ; Humans ; Lymphoma ; metabolism ; pathology ; Receptors, Notch ; metabolism ; Signal Transduction

OBJECTIVETo observe the effects of lead on mRNA and protein expression of PKC in U251 cell line.

METHODSAfter U251 cells were exposed to 0.05, 0.50, 5.00, 50.00, 500.00, 900.00 and 1000.00 micromol/L Ph(Ac)2 for 24 hours, the cytotoxicity of Pb on U251 cells was measured by MTT assay. RT-PCR and Western blot assay were used to detect the mRNA and protein expression levels of PKC in U251 cells exposed to 0.05, 5.00 and 500.00 micromol/L Ph (Ac), for 24 hours.

RESULTSThe survival rates of U251 cells treated with 5.00, 50.00, 500.00, 900.00 and 1000.00 micromol/L Pb (Ac)2 were 84.5%, 78.2%, 76.5%, 50.3% and 43.2%, respectively, which were significantly lower than those of control group (P < 0.01). The PKC mRNA expression level (0.40 +/- 0.01) of U251 cells treated with 500.00 micromol/L Pb (Ac)2 was significantly lower than that (0.51 +/- 0.02) of control group (P < 0.01). The PKC protein expression levels of U251 cells treated with 0.05, 5.00 or 500.00 micromol/L Pb(Ac)2 were 0.68 +/- 0.02, 0.62 +/- 0.01 and 0.33 +/- 0.02, respectively, which were significantly lower (0.98 +/- 0.01) than those of control group (P < 0.01).

CONCLUSIONLead can decline the cell viability, PKC mRNA and protein expression levels of U251 cells.

Cell Line, Tumor ; Cell Survival ; Humans ; Lead ; toxicity ; Protein Kinase C ; metabolism ; RNA, Messenger ; metabolism

Patients with glioblastoma (GBM) generally have a bad prognosis and short overall survival after being treated with surgery, chemotherapy or radiotherapy due to the histological heterogeneity, strong invasive ability and rapid postoperative recurrence of GBM. The components of GBM cell-derived exosome (GBM-exo) can regulate the proliferation and migration of GBM cell via cytokines, miRNAs, DNA molecules and proteins, promote the angiogenesis via angiogenic proteins and non-coding RNAs, mediate tumor immune evasion by targeting immune checkpoints with regulatory factors, proteins and drugs, and reduce drug resistance of GBM cells through non-coding RNAs. GBM-exo is expected to be an important target for the personalized treatment of GBM and a marker for diagnosis and prognosis of this kind of disease. This review summarizes the preparation methods, biological characteristics, functions and molecular mechanisms of GBM-exo on cell proliferation, angiogenesis, immune evasion and drug resistance of GBM to facilitate developing new strategies for the diagnosis and treatment of GBM.
Humans ; Glioblastoma/genetics* ; Exosomes/metabolism* ; MicroRNAs/metabolism* ; Prognosis ; Cell Proliferation ; Brain Neoplasms/genetics* ; Cell Line, Tumor

OBJECTIVETo search for a new diagnostic biomarker for prostate cancer by comparing the differences in the expressions of netrin-1 and UNC5B in prostate cancer cells with different invasive abilities.

METHODSWe examined the expressions of netrin-1 and UNC5B in five prostate cancer cell lines DU145, 22RV1, PC3, PC3M and RWPE-1 using RT-PCR and Western blot, and positioned the ligands netrin-1 and its receptor UNC5B in the prostate cancer cells by immunofluorescence.

RESULTSBoth netrin-1 and UNC5B were expressed in the prostate cancer cells, and the expression of netrin-1 was significantly increased in highly invasive cells (P < 0.05), while that of UNC5B in RWPE-1 (normal) cells (P < 0.05).

CONCLUSIONThe expressions of netrin-1 and UNC5B are closely related to the infiltration and progression of prostate cancer, and expected to be as potential biomarkers for predicting the malignancy degree of prostate cancer.

Biomarkers, Tumor ; metabolism ; Cell Line, Tumor ; Humans ; Male ; Nerve Growth Factors ; metabolism ; Netrin-1 ; Prostatic Neoplasms ; metabolism ; pathology ; Receptors, Cell Surface ; metabolism ; Tumor Suppressor Proteins ; metabolism

Cell Differentiation ; Cell Line, Tumor ; metabolism ; Cell Proliferation ; Humans ; Trans-Activators ; biosynthesis ; genetics ; Transfection