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1.
BMC Musculoskelet Disord ; 25(1): 337, 2024 Apr 26.
Article in English | MEDLINE | ID: mdl-38671386

ABSTRACT

PURPOSE: This study aimed to compare cervical sagittal parameters and clinical outcomes between patients undergoing cervical laminoplasty(CL) and those undergoing lateral mass screw fixation(LMS). METHODS: We retrospectively studied 67 patients with multilevel ossification of the posterior longitudinal ligament (OPLL) of the cervical spine who underwent lateral mass screw fixation (LMS = 36) and cervical laminoplasty (CL = 31). We analyzed cervical sagittal parameters (C2-7 sagittal vertical axis (C2-7 SVA), C0-2 Cobb angle, C2-7 Cobb angle, C7 slope (C7s), T1 slope (T1s), and spino-cranial angle (SCA)) and clinical outcomes (visual analog scale [VAS], neck disability index [NDI], Japanese Orthopaedic Association [JOA] scores, recovery rate (RR), and minimum clinically significant difference [MCID]). The cervical sagittal parameters at the last follow-up were analyzed by binary logistic regression. Finally, we analyzed the correlation between the cervical sagittal parameters and each clinical outcome at the last follow-up after surgery in both groups. RESULTS: At the follow-up after posterior decompression in both groups, the mean values of C2-C7 SVA, C7s, and T1s in the LMS group were more significant than those in the CL group (P ≤ 0.05). Compared with the preoperative period, C2-C7 SVA, T1s, and SCA gradually increased, and the C2-C7 Cobb angle gradually decreased after surgery (P < 0.05). The improvement in the JOA score and the recovery rate was similar between the two groups, while the improvement in the VAS-N score and NDI score was more significant in the CL group (P = 0.001; P = 0.043). More patients reached MCID in the CL group than in the LMS group (P = 0.036). Binary logistic regression analysis showed that SCA was independently associated with whether patients reached MCID at NDI postoperatively. SCA was positively correlated with cervical NDI and negatively correlated with cervical JOA score at postoperative follow-up in both groups (P < 0.05); C2-7 Cobb angle was negatively correlated with cervical JOA score at postoperative follow-up (P < 0.05). CONCLUSION: CL may be superior to LMS in treating cervical spondylotic myelopathy caused by OPLL. In addition, smaller cervical SCA after posterior decompression may suggest better postoperative outcomes.


Subject(s)
Bone Screws , Cervical Vertebrae , Laminoplasty , Ossification of Posterior Longitudinal Ligament , Humans , Ossification of Posterior Longitudinal Ligament/surgery , Ossification of Posterior Longitudinal Ligament/diagnostic imaging , Laminoplasty/methods , Female , Male , Cervical Vertebrae/surgery , Cervical Vertebrae/diagnostic imaging , Retrospective Studies , Middle Aged , Aged , Treatment Outcome , Decompression, Surgical/methods , Follow-Up Studies
2.
J Orthop Surg Res ; 19(1): 264, 2024 Apr 25.
Article in English | MEDLINE | ID: mdl-38664852

ABSTRACT

OBJECTIVE: This study aimed to evaluate the influence of herniation of cartilaginous endplates on postoperative pain and functional recovery in patients undergoing percutaneous endoscopic lumbar discectomy (PELD) for lumbar disc herniation (LDH). METHODS: A retrospective analysis was conducted on 126 patients with LDH treated with PELD at the Third Hospital of Hebei Medical University from January 2021 to January 2022. Whether cartilaginous endplates had herniated was identified by analyzing these specific findings from MRI scans: posterior marginal nodes, posterior osteophytes, mid endplate irregularities, heterogeneous low signal intensity of extruded material, and Modic changes in posterior corners and mid endplates. Patients were assessed for postoperative pain using the Visual Analogue Scale (VAS) and functional recovery using the Oswestry Disability Index (ODI) and Modified MacNab criteria. Statistical analyses compared outcomes based on the presence of herniation of cartilaginous endplates. RESULTS: Patients with herniation of cartilaginous endplates experienced higher pain scores early postoperatively but showed significant improvement in pain and functional status over the long term. The back pain VAS scores showed significant differences between the groups with and without herniation of cartilaginous endplates on postoperative day 1 and 1 month (P < 0.05). Leg pain VAS scores showed significant differences on postoperative day 1 (P < 0.05). Modic changes were significantly associated with variations in postoperative recovery, highlighting their importance in predicting patient outcomes. In patients with herniation of cartilaginous endplates, there were statistically significant differences in the back pain VAS scores at 1 month postoperatively and the ODI functional scores on postoperative day 1 between the groups with and without Modic changes (P < 0.05). There were no significant differences in the surgical outcomes between patients with and without these conditions regarding the Modified MacNab criteria (P > 0.05). CONCLUSION: Herniation of cartilaginous endplates significantly affect early postoperative pain and functional recovery in LDH patients undergoing PELD. These findings emphasize the need for clinical consideration of these imaging features in the preoperative planning and postoperative management to enhance patient outcomes and satisfaction.


Subject(s)
Diskectomy, Percutaneous , Endoscopy , Intervertebral Disc Displacement , Lumbar Vertebrae , Recovery of Function , Humans , Intervertebral Disc Displacement/surgery , Intervertebral Disc Displacement/diagnostic imaging , Male , Female , Diskectomy, Percutaneous/methods , Retrospective Studies , Lumbar Vertebrae/surgery , Lumbar Vertebrae/diagnostic imaging , Middle Aged , Adult , Endoscopy/methods , Pain, Postoperative/etiology , Treatment Outcome , Pain Measurement , Cartilage/diagnostic imaging , Aged , Magnetic Resonance Imaging
3.
Orthop Surg ; 16(2): 329-336, 2024 Feb.
Article in English | MEDLINE | ID: mdl-38093558

ABSTRACT

OBJECTIVE: Previous studies have shown that cervical sagittal alignment is strongly associated with cervical deformity, myelopathy, and cervical adjacent-segmental disease, and these cervical sagittal parameters are correlated with health-related quality of life. However, less attention has been paid to cervical sagittal balance in various cervical disorders. This study aimed to compare cervical sagittal parameters between patients with nonspecific neck pain (NS-NP) and patients with cervical spondylotic radiculopathy (CSR) and cervical spondylotic myelopathy (CSM). METHODS: We retrospectively examined 236 patients from between January 2020 and October 2022. We divided them into three groups (NS-NP, CSR, and CSM) and collected general information and cervical sagittal parameters for these patients. The variation of parameters between the size of these parameters and gender differences was analyzed. Pearson's or Spearman's correlation was applied to analyze the association of cervical sagittal parameters of all patients between the three groups. RESULTS: There were significant differences in age and sex among the three groups (p < 0.001), with the NS-NP group being the youngest and NS-NP being more common in women. The parameters of cervical sagittal position significantly differed among the three groups (p < 0.05). Pearson's or Spearman's correlation result showed that the C2-C7 Cobb angle was negatively associated with the C2-C7 sagittal vertical angle (SVA), and the C2-C7 Cobb angle and T1 slope (T1s) were negatively associated with the spino-cranial angle (SCA). There was a positive correlation between the C2-C7 Cobb angle and C7 slope (C7s), C2-C7 SVA and T1s, C2-C7 SVA and SCA, and C7s and T1s. CONCLUSION: This study showed that between the three groups, patients with nonspecific neck pain had smaller SCA, and among patients with NS-NP, women had more significant SCA. The smaller anteroposterior diameter of the thorax in women might explain this difference.


Subject(s)
Lordosis , Radiculopathy , Spinal Cord Diseases , Spondylosis , Humans , Female , Neck Pain/etiology , Radiculopathy/complications , Quality of Life , Retrospective Studies , Cervical Vertebrae/diagnostic imaging , Spondylosis/complications , Spondylosis/diagnostic imaging , Spinal Cord Diseases/diagnostic imaging
4.
Front Surg ; 10: 1121807, 2023.
Article in English | MEDLINE | ID: mdl-37091266

ABSTRACT

Objective: The purpose of this study was to introduce enhanced recovery after surgery (ERAS) concept into patients with lumbar degenerative diseases who were treated with oblique lumbar interbody fusion (OLIF), and to assess whether it could increase clinical efficacy, reduce perioperative complications, shorten length of hospital stay (LHS), decrease readmission rate, and improve patient satisfaction. Methods: The study included patients with lumbar degenerative diseases (LDDs) who underwent OLIF between July 2017 and October 2018 (non-ERAS group), and between November 2018 and July 2020 (ERAS group). The two groups were compared according to the demographic and clinical characteristics. Results: There was no significant difference in descriptive characteristics and concomitant diseases between the two groups. The preoperative Oswestry disability index (ODI) score (P = 0.191), lumbar visual analogue scale (VAS) score (P = 0.470), and leg VAS score (P = 0.657) did not significantly different. Most of the ERAS measures were also well implemented after surgery, except for early delivery (74.2%), early catheter removal (63.9%), and multimodal analgesia (80.6%). The LHS in the ERAS group was significantly shorter than that in the non-ERAS group (P = 0.004). Besides, Hamilton Anxiety Rating Scale (HAMA) score at 3 days after surgery showed a significant difference between the two groups (P = 0.019). The patient satisfaction in ERAS group was significantly higher than that in the non-ERAS group (P = 0.001). Conclusion: The new nursing pattern combined with ERAS in patients with LDDs who underwent OLIF did not improve the short-term prognosis of surgery, while it could effectively reduce postoperative complications, shorten the LHS, and improve patient satisfaction, and did not lead to additional adverse events.

5.
Exp Brain Res ; 239(8): 2435-2444, 2021 Aug.
Article in English | MEDLINE | ID: mdl-34106297

ABSTRACT

Hypothyroidism causes somatic, psychosocial and affective psychosis, including depression-like behaviors. In this study, (hypothyroidism group; HP group) adult male Sprague Dawley (SD) rats were induced to hypothyroidism after 5 weeks of exposure to 0.05% propylthiouracil (PTU) in potable water, control animals (CON group) were given the same amount of water. The following behavioral experiments were conducted, respectively: open-field test (OFT), forced swimming test (FST), tail suspension test (TST). TT[Formula: see text] and TT[Formula: see text] levels were measured after the behavior tests and the expression levels of 5-HT[Formula: see text] receptor and 5-HT[Formula: see text] receptor proteins were analyzed in the hippocampus and prefrontal cortex. The level of TT[Formula: see text] and TT[Formula: see text] in the HP group rats was much lower than that in the CON group. The hypothyroid rats also showed weight loss, much longer immobility time in tail suspension test and forced swimming test. Besides, 5 weeks of PTU administration was associated with significantly decreased expression levels of 5-HT[Formula: see text] receptor and 5-HT[Formula: see text] receptor proteins compared with control group, which were significantly negatively correlated with immobility time in FST and TST. In conclusion, our results suggest that hypothyroidism induces depressive behaviors through the influence of the serotonin system, and the decreased expression of the 5-HT[Formula: see text] receptor is an important cause of the depressive behaviors in hypothyroidism.


Subject(s)
Depression , Hypothyroidism , Animals , Behavior, Animal , Depression/etiology , Hypothyroidism/chemically induced , Hypothyroidism/complications , Male , Rats , Rats, Sprague-Dawley , Receptor, Serotonin, 5-HT2A , Serotonin , Swimming
6.
Curr Drug Metab ; 21(7): 541-547, 2020.
Article in English | MEDLINE | ID: mdl-32586251

ABSTRACT

BACKGROUND: The liver is one of the major organ involved in drug metabolism. Cytochrome P450s are predominantly involved in drug metabolism. A wide range of CYPs have been reported in the liver which have been involved in its normal as well as in diseased conditions. Doxorubicin, one of the most potent chemotherapeutic drugs, although highly efficacious, also has adverse side effects, with its targets being liver and cardiac tissue. OBJECTIVE: The study aims to evaluate the reversal potentials of berberine on Doxorubicin induced cyp conversion. METHODOLOGY: In the present study, the interplay between anti-oxidants, cytochrome and inflammatory markers in DOX induced liver toxicity and its possible reversal by berberine was ascertained. RESULTS: DOX administration significantly elevated serum as well as tissue stress, which was reverted by berberine treatment. A similar response was observed in tissue inflammatory mediators as well as in serum cytokine levels. Most profound reduction in the cytochrome expression was found in Cyp 2B1, 2B2, and 2E1. However, 2C1, 2C6, and 3A1 although showed a decline, but it did not revert the expression back to control levels. CONCLUSION: It could be concluded that berberine may be an efficient anti-oxidant and immune modulator. It possesses low to moderate cytochrome modulatory potentials.


Subject(s)
Antibiotics, Antineoplastic , Antioxidants/therapeutic use , Berberine/therapeutic use , Chemical and Drug Induced Liver Injury/drug therapy , Cytochrome P-450 Enzyme System/metabolism , Doxorubicin , Immunologic Factors/therapeutic use , Animals , Antioxidants/pharmacology , Berberine/pharmacology , Chemical and Drug Induced Liver Injury/immunology , Chemical and Drug Induced Liver Injury/metabolism , Chemical and Drug Induced Liver Injury/pathology , Cytokines/blood , Immunologic Factors/pharmacology , Liver/drug effects , Liver/metabolism , Liver/pathology , Male , NF-kappa B/genetics , Rats, Wistar , Toll-Like Receptor 2/genetics , Toll-Like Receptor 4/genetics
7.
Onco Targets Ther ; 8: 2271-7, 2015.
Article in English | MEDLINE | ID: mdl-26345235

ABSTRACT

In the present study, we analyzed microRNA (miRNA) and gene expression profiles using 499 papillary thyroid carcinoma (PTC) samples and 58 normal thyroid tissues obtained from The Cancer Genome Atlas database. A pivotal regulatory network of 18 miRNA and 16 targets was identified. Upregulated miRNAs (miR-222, miR-221, miR-146b, miR-181a/b/d, miR-34a, and miR-424) and downregulated miRNAs (miR-9-1, miR-138, miR-363, miR-20b, miR-195, and miR-152) were identified. Among them, the upregulation of miR-424 and downregulation of miR-363, miR-195, and miR-152 were not previously identified. The genes CCNE2 (also known as cyclin E2), E2F1, RARA, CCND1 (cyclin D1), RUNX1, ITGA2, MET, CDKN1A (p21), and COL4A1 were overexpressed, and AXIN2, TRAF6, BCL2, RARB, HSP90B1, FGF7, and PDGFRA were downregulated. Among them, CCNE2, COL4A1, TRAF6, and HSP90B1 were newly identified. Based on receiver operating characteristic curves, several miRNAs (miR-222, miR-221, and miR-34a) and genes (CCND1 and MET) were ideal diagnostic indicators, with sensitivities and specificities greater than 90%. The combination of inversely expressed miRNAs and targets improved diagnostic accuracy. In a clinical feature analysis, several miRNAs (miR-34a, miR-424, miR-20b, and miR-152) and genes (CCNE2, COL4A1, TRAF6, and HSP90B1) were associated with aggressive clinical features, which have not previously been reported. Our study not only identified a pivotal miRNA regulatory network associated with PTC but also provided evidence that miRNAs and target genes can be used as biomarkers in PTC diagnosis and clinical risk evaluation.

8.
Cell Physiol Biochem ; 36(6): 2466-79, 2015.
Article in English | MEDLINE | ID: mdl-26279448

ABSTRACT

BACKGROUND/AIMS: salusin-ß is considered to be a potential pro-atherosclerotic factor. Regulation and function of vascular smooth muscle cells (VSMCs) are important in the progression of atherosclerosis. Peroxisome proliferator-activated receptor gamma (PPARγ) exerts a vascular protective role beyond its metabolic effects. Salusin-ß has direct effects on VSMCs. The aim of the present study was to assess the effect of salusin-ß on PPARγ gene expression in primary cultured rat VSMCs. METHODS: Western blotting analysis, real-time PCR and transient transfection approach were used to determine expression of target proteins. Specific protein knockdown was performed with siRNA transfection. Cell proliferation was determined by 5-bromo-2'-deoxyuridine incorporation. The levels of inflammation indicators interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) were determined using enzyme-linked immunosorbent assay. RESULTS: Salusin-ß negatively regulated PPARγ gene expression at protein, mRNA and gene promoter level in VSMCs. The inhibitory effect of salusin-ß on PPARγ gene expression contributed to salusin-ß-induced VSMCs proliferation and inflammation in vitro. IκBα-NF-κB activation, but not NF-κB p50 or p65, mediated the salusin-ß-induced inhibition of PPARγ gene expression. Salusin-ß induced nuclear translocation of histone deacetylase 3 (HDAC3). HDAC3 siRNA prevented salusin-ß-induced PPARγ reduction. Nuclear translocation of HDAC3 in response to salusin-ß was significantly reversed by an IκBα inhibitor BAY 11-7085. Furthermore, IκBα-HDAC3 complex was present in the cytosol of VSMCs but interrupted after salusin-ß treatment. CONCLUSION: IκBα-HDAC3 pathway may contribute to salusin-ß-induced inhibition of PPARγ gene expression in VSMCs.


Subject(s)
Gene Expression Regulation/drug effects , Intercellular Signaling Peptides and Proteins/pharmacology , Muscle, Smooth, Vascular/cytology , Myocytes, Smooth Muscle/metabolism , PPAR gamma/genetics , Animals , Cell Nucleus/drug effects , Cell Nucleus/metabolism , Cell Proliferation/drug effects , Down-Regulation/drug effects , Histone Deacetylase Inhibitors/pharmacology , Histone Deacetylases/metabolism , I-kappa B Proteins/metabolism , Inflammation/genetics , Inflammation/pathology , Ligands , Male , Myocytes, Smooth Muscle/drug effects , NF-KappaB Inhibitor alpha , NF-kappa B/metabolism , PPAR gamma/metabolism , Prostaglandin D2/analogs & derivatives , Prostaglandin D2/pharmacology , Protein Transport/drug effects , RNA, Small Interfering/metabolism , Rats, Sprague-Dawley
9.
Int J Mol Med ; 36(4): 1097-103, 2015 Oct.
Article in English | MEDLINE | ID: mdl-26252081

ABSTRACT

Thyroid carcinoma (TC) is the most common malignancy of the endocrine system, and papillary thyroid carcinoma (PTC) accounts for the largest proportion of cases with TC. Although histology is considered the gold standard in the diagnosis of PTC, the sensitivity and specificity of this method is low. Therefore, developing novel diagnostic and prognostic biomarkers for PTC is essential. MicroRNAs (miRNAs or miRs) and their target RNAs play critical roles in tumorigenesis and tumor progression. Thus, the characteristic miRNA and mRNA expression profiles may function as diagnostic biomarkers for tumors, making it possible to predict the tumor stage and the prognosis of patients. In the present study, we detected miRNAs and mRNAs which can function as novel biomarkers for the diagnosis of PTC. The sensitivity of the diagnostic tests was evaluated by receiver operating characteristic curve analysis. Pearson's correlation analysis was used to determine the correlation between mRNAs and miRNAs, and cancer types. We found that the area under the curve (AUC) values of 8 miRNAs (miR-106a, miR-15a, miR-30a, miR-30b, miR-20a, miR-20b, miR-30d and miR-30e) and 8 mRNAs [axis inhibition protein 2 (AXIN2), integrin, alpha 3 (antigen CD49C, alpha 3 subunit of VLA-3 receptor) (ITGA3), tumor protein p53 inducible nuclear protein (TP53INP)1, TP53INP2, B-cell CLL/lymphoma 2 (BCL2), phosphatase and tensin homolog (PTEN), FOS and K(lysine) acetyltransferase 2B (KAT2B)] were >0.90. The combination of miR-15a and AXIN2 significantly improved the diagnostic accuracy. Therefore, our data indicate that the differential expression of miRNAs combined with that of their target mRNAs may serve as a powerful biomarker for distinguishing PTC from benign tissues.


Subject(s)
Biomarkers, Tumor/biosynthesis , Carcinoma/diagnosis , Carcinoma/metabolism , MicroRNAs/biosynthesis , Neoplasm Proteins/biosynthesis , RNA, Messenger/biosynthesis , RNA, Neoplasm/biosynthesis , Thyroid Neoplasms/diagnosis , Thyroid Neoplasms/metabolism , Biomarkers, Tumor/genetics , Carcinoma/genetics , Carcinoma, Papillary , Female , Humans , Male , MicroRNAs/genetics , Neoplasm Proteins/genetics , RNA, Messenger/genetics , RNA, Neoplasm/genetics , Thyroid Cancer, Papillary , Thyroid Neoplasms/genetics
10.
J Cardiovasc Pharmacol ; 65(4): 377-85, 2015 Apr.
Article in English | MEDLINE | ID: mdl-25551321

ABSTRACT

Despite the clear mitogenic effect of salusin-ß on vascular smooth muscle cells (VSMCs), which contributes to its proatherosclerotic effects, additional studies are needed to explore its underlying mechanisms. The aim of this study was to investigate the mechanism of salusin-ß's effects on VSMCs cell cycle regulation and the possible signal pathways. Salusin-ß accelerated the G1/S phase transition in VSMCs and increased the expression levels of cyclins D1 and E. Silencing either cyclin D1 or cyclin E gene inhibited salusin-ß-induced VSMCs proliferation, cell cycle progression, phosphorylation of the Rb protein, and dissociation of the E2F-Rb complex. Importantly, expression of cyclin E was also induced by cyclin D1. Next, we found that salusin-ß increased the protein expressions of activator protein-1 (AP-1) subunits c-Jun and c-Fos, and enhanced binding of AP-1 to the promoter region of cyclin D1. In addition, inhibition of AP-1 activity could lead to significant suppression of salusin-ß-induced cyclin D1 expression. Furthermore, MPAKs pathways were found to mediate salusin-ß-induced VSMCs proliferation, cyclin D1, cyclin E, c-Jun, and c-Fos expression. These results suggest that salusin-ß promotes cell cycle progression of VSMCs by upregulating the cyclin D1 and cyclin E, in an AP-1-dependent manner through mitogen-activated protein kinases signaling pathways.


Subject(s)
Cell Proliferation/physiology , Intercellular Signaling Peptides and Proteins/metabolism , Muscle, Smooth, Vascular/physiology , Myocytes, Smooth Muscle/physiology , Animals , Cell Cycle/physiology , Cells, Cultured , Cyclin D1/metabolism , Cyclin E/metabolism , Maturation-Promoting Factor/metabolism , Rats , Signal Transduction
12.
Oncol Rep ; 29(4): 1415-20, 2013 Apr.
Article in English | MEDLINE | ID: mdl-23380809

ABSTRACT

The incidence of thyroid cancer has recently experienced a rapid increase in China, and papillary thyroid carcinoma (PTC) accounts for nearly 80% of human thyroid cancers. In the present study, the differential expression of microRNAs (miRNAs) and their target genes were identified in order to analyze the potential roles of miRNAs as biomarkers and in papillary thyroid carcinogenesis. One hundred and twenty-six PTC samples were collected from patients at the China-Japan Union Hospital, China, and the gene/miRNA expression profiles were examined with Illumina BeadChips and verified by real­time RT-PCR. Gene Ontology (GO) categories were determined, and pathway analysis was carried out using KEGG. miRNA target genes were predicted by implementing three computational analysis programs: TargetScanS, DIANA-microT and PicTar. Two hundred and forty-eight miRNAs and 3,631 genes were found to be significantly deregulated (gene, P<0.05; miRNA, P<0.01) in PTC tissues when compared with their matching normal thyroid tissues. hsa-miR-206 (target gene, MET), hsa-miR-299-3p (target gene, ITGAV), hsa-miR-101 (target gene, ITGA3), hsa-miR-103 (target gene, ITGA2), hsa-miR­222 (target genes, KIT and AXIN2), hsa-miR-15a (target genes, AXIN2 and FOXO1) and hsa-mir-221 (target gene, KIT) were identified. Together with the functions of the target genes, we further elucidated the role of miRNAs in papillary thyroid carcinogenesis and suggest the use of miRNAs as biomarkers for early diagnosis. Our findings provide the basis for future studies in the field of miRNA-based cancer therapy.


Subject(s)
Carcinoma/genetics , Gene Regulatory Networks , MicroRNAs/genetics , Molecular Targeted Therapy , Thyroid Neoplasms/genetics , Carcinoma/pathology , Carcinoma, Papillary , Gene Expression Profiling , Gene Expression Regulation, Neoplastic , Humans , Oligonucleotide Array Sequence Analysis , Thyroid Cancer, Papillary , Thyroid Neoplasms/pathology
13.
Cancer Biother Radiopharm ; 28(2): 153-9, 2013 Mar.
Article in English | MEDLINE | ID: mdl-23268708

ABSTRACT

PURPOSE: Cell death is one of the most important endpoints of radiosensitivity. The tumor suppressor p53 participates not only in regulation of apoptosis, but also in autophagy mechanism. In this study, H1299-P53 (with wild-type p53) and H1299-175H (with mutant 175H) were used, and the effects of p53 on radiosensitivity were analyzed. METHODS: Cell models with different p53 status were established by gene engineering, and cell viability was examined by colony formation assay, and cell counting kit-8 (CCK-8), 3-Methyladenine, and Z-VAD were used to block autophagy and apoptosis, respectively. Western blot was used to detect protein expression; monodansylcadaverine (MDC) staining was used to analyze autophagy rate; DAPI/Propidium Iodide (PI) staining and flow cytometry were used to assess apoptosis and necrosis. RESULTS: In parental H1299, H1299-P53, and H1299-175H cells, radiosensitivity exhibited different by colony formation and CCK-8 assay (D0: 1.764 Gy, 1.407 Gy and 1.695 Gy; Dq: 2.977 Gy, 1.199 Gy and 2.312 Gy in turn). The radiosensitization of p53 was associated with the increase of MDM2 and P21 expression. The ionizing radiation (IR)-induced apoptosis was significant in H1299-P53 compared with in H1299 and H199-175H (p < 0.05) by flow cytometry, and the expression of cleaved-caspase3 was increased in H1299-P53 cells. While the IR-induced autophagy was significant in H1299 cells (p < 0.01) and decreased in H1299-P53 and H1299-175H cells (p < 0.01) by MDC staining, the expression of MAPLC3II and Beclin-1 increased in H1299, but not in H1299-p53 and H199-175H cells. The IR-induced cell survival was significantly increased by Z-VAD-FMK and decreased by 3MA in H1299-P53 cells; IR- induced autophagy was significantly increased by Z-VAD-FMK in H1299-P53 cells (p < 0.01), but not changed in H1299 cells. CONCLUSION: p53 could regulate radiosensitivity by inhibiting autophagy and activating apoptosis; autophagy provides a prosurvival mechanism, and p53 potently abrogated the IR-induced autophagy, while mutant 175H shown no effect on radiosensitivity, suggesting that individual treatment strategies should be based on p53 status in patients.


Subject(s)
Apoptosis/radiation effects , Autophagy/radiation effects , Lung Neoplasms/pathology , Radiation Tolerance/physiology , Tumor Suppressor Protein p53/genetics , Tumor Suppressor Protein p53/metabolism , Blotting, Western , Colony-Forming Units Assay , Humans , Lung Neoplasms/metabolism , Lung Neoplasms/radiotherapy , Transcriptional Activation , Tumor Cells, Cultured , X-Rays
14.
Radiat Oncol ; 7: 213, 2012 Dec 17.
Article in English | MEDLINE | ID: mdl-23244773

ABSTRACT

PURPOSE: Autophagy has attracted attentions as a novel mechanism for tumor development. In this study Human ovarian carcinoma cell line SKOV3 and multidrug-resistant phenotype SKVCR cells were used and the roles of autophagy in radiation-induced cell death were analyzed. METHODS AND MATERIALS: Cell viability was examined by colony formation and cell counting kit-8 (CCK-8) assay, 3MA and ZVAD were used to block autophagy and apoptosis, respectively. Quantitative real-time PCR was used to detect mRNA level and Western blot was used to detect protein expression, monodansylcadaverine (MDC) staining and flow cytometery were used for autophagy, apoptosis and cell cycle dynamics, respectively. RESULTS: (1) The radiosensitivity exhibited differently in SKOV3 and SKVCR cells (SKOV3: D0=3.37, SKVCR: D0= 4.18); compared with SKOV3 the constitutive expression of MAPLC3 in SKVCR was higher, but no change of Caspase-3 and cleaved Caspase-3. (2) The ionizing radiation (IR)- induced apoptosis and autophagy were significant in both cells (P<0.05); inhibition of apoptosis with ZVAD showed no impact on survival of SKOV3 and SKVCR cells after radiation, while inhibition of autophagy significantly decreased viability in SKVCR cells, for SKVO3 cells only low level of radiation (2 Gy and 4 Gy) could decrease the viability(P<0.05). (3) ZVAD inhibited apoptosis and autophagy in both cells, 3MA inhibit apoptosis in SKOV3, and promote apoptosis in SKVCR, together with inhibition of autophagy. (4) G2/M arrest was induced by radiation in both cells; the accumulation of G2/M was more significant in SKOV3, 3MA attenuated the radiation-induced S phase delay in SKVCR. CONCLUSION: IR-induced autophagy provides a self-protective mechanism against radiotherapy in SKVCR cells, the use of autophagy inhibitor, 3MA, increases the killing effects of radiation by inhibiting autophagy and radiation- induced S phase delay, also by the increase of apoptosis, which suggests a better therapeutic strategy in drug- resistant SKVCR ovarian cancer cells.


Subject(s)
Autophagy/physiology , Ovarian Neoplasms/radiotherapy , Apoptosis/radiation effects , Caspase 3/genetics , Cell Cycle Checkpoints/radiation effects , Cell Line, Tumor , Drug Resistance, Multiple , Drug Resistance, Neoplasm , Female , Humans , Microtubule-Associated Proteins/genetics , Oligopeptides/pharmacology , Ovarian Neoplasms/drug therapy , Ovarian Neoplasms/pathology , Radiation Tolerance
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