Effect of YAP1 on erlotinib-resistance in lung adenocarcinoma cells / 军事医学

Objective To explore the effect of Yes-associated protein 1 (YAP1) and potential mechanism on erlotinib ( ER) resistance in lung adenocarcinoma.Methods In PC-9 cells and acquired ER resistant PC-9 ( PC-9/ER) cells, the expression changes in YAP1 gene were measured by quantitative real-time PCR( RT-qPCR) and Western blot.Indirect immunofluorescence was adopted to observe the location of YAP1.PC-9/ER cells were treated with the verteporfin ( VP, YAP1 inhibitor) for 24 h and 48 h, respectively.Expression changes in mRNA and proteins of YAP1, AKT and p-AKT were detected in the presence or absence of VP.The effect of VP was analyzed by drug resistance index using Cell Counting Kit 8(CCK-8) assay.Results The resistance index of PC-9/ER cells was (99.80 ±25.81).Compared with PC-9 cells, the expression levels of YAP1 mRNA and protein were increased in PC-9/ER.The inhibitory efficiency of VP was (50.96 ±5.86)%, and the levels of AKT and p-AKT proteins were down-regulated by the inhibition of YAP1 simultaneously.The half maximal inhibitory concentration (IC50) of PC-9/ER decreased from (11.10 ±2.72) to (1.47 ± 0.32)μmol/L (P =0.024).Resistance index was reduced to one eighth of the original.Conclusion These results indicate that the YAP1 mediates ER resistance in lung adenocarcinoma.Suppression of YAP1 can reduce the resistance through PI3K/AKT signaling pathway.Therefore, YAP1 may be a potential target for lung cancer gene therapy.

The value of detecting plasma microRNA-125a-3p,IGF-2 on monitoring invasion and metastasis in NSCLC / 重庆医学

Objective To investigate the value of detecting plasma microRNA‐125a‐3p(miRNA‐125a‐3p) ,IGF‐2 on monitoring invasion and metastasis in NSCLC ,and to study the correlation between miR‐125a‐3p and IGF‐2 .Methods miR‐125a‐3p transcripts of 20 controls ,73 NSCLC were performed in plasma by quantitative reverse transcription‐polymerase chain reaction(qRT‐PCR) and PCR data was analyzed by the 2‐ΔΔCT method .The expression of IGF‐2 in plasma was detected by ELISA .Results The expression of miR‐125a‐3p in stage Ⅲ /Ⅳ was lower than stage Ⅰ/Ⅱ and the controls(P=0 .001 ,P=0 .005) .There was no statistical differ‐ence between the stage Ⅰ /Ⅱ patients and the controls(P=0 .776) .The expression of miR‐125a‐3p was related with lymph node metastas ,lower expression in positive lymph node metastasis (P=0 .003) .The expression of IGF‐2 in stage Ⅰ /Ⅱ 、stage Ⅲ /Ⅳ was higher than the controls(P=0 .036 ,P=0 .011) .There was no statistical difference between the stageⅠ/Ⅱ and stage Ⅲ/Ⅳ (P=0 .451) . The expression of IGF‐2 was related with lymph node metastas ,higher expression in positive lymph node metastasis (P=0 .037) .The re‐sults showed a negative correlation between miR‐125a‐3p expression and IGF‐2 in plasma(r= -0 .280 ,P=0 .007) .Conclusion Low ex‐pression of miR‐125a‐3p and high expression of IGF‐2 in plasma may play a role in invasion and metastasis of NSCLC .miR‐125a‐3p may play a negative regulatory role on IGF‐2 .

Design and application of computer-assisted cancer chemotherapy plan and management system / 中华医学教育探索杂志

Chemotherapy treatment plays an important role in the comprehensive treatment of malignant disease,but the chemotherapy related knowledge and the selection of an appropriate regimen for certain patient are hard to master.The establishment of computer-assisted cancer chemotherapy program and management system with a follow-up database of cancer patients can help the oncologist to master the designing skill of chemotherapy regimen and cancer-related knowledge quickly,improve the teaching qnality and the efficiency of treating malignant diseases.

Expression of membrane inhibitor of reactive lysis in non-small cell lung carcinoma and its clinical significance / 中国基层医药

Objective To survey membrane inhibitor of reactive lysis(MIRL) expression in non-small cell lung careinoma(NSCLC) and to analyze the relationship between MIRL expression and clinical staging, adjuvant chemotherapy and disease-free survial. Methods The expression of MIRL in 8 adjacent tissues and 36 NSCLC sam-pies were determined by immunohistochemistry. Furthermore, the relationship between MIRL expression and clinical stage ,adjuvant chemotherapy and disease-free survival was assayed by follow-up. Results Among 36 samples of non-small-cell lung cancer,there were 10(27.8%) samples expressing MIRE. Out of 18 samples of squamous carcinoma, 4(22.2%) expressed MIRL,while 6(37.5%) expressed it in 16 samples of adenocarcinoma,there was no statistical significance between them(P>0.05). There were no expression in 2 samples of large cell carcinoma. There was no correlation between MIRL expression and disease-free survival(P>0.05). MIRL positive expression rate in patients with preoperational adjuvant chemotherapy was significantly lower than that of those without preoperational adjuvant chemotherapy(P<0.05). Conclusions There is great percentage of MIRE expression in NSCLC. Our present study suggests that the immunological inhibition of MIRL should be blocked when monoclonal antibody is used in the treat-merit of NSCLC.

Progress in research of genetic circuits / 生物医学工程学杂志

Genetic circuits are collections of basic elements that interact to produce a particular behavior. By constructing biochemical logic circuits and embedding them in cells, one can extend or modify the behavior of cells. To date, several small synthetic gene networks have been built that accomplish specific genetic regulatory functions in vivo: the autorepressor, in which a repressor regulates its own production to reduce noise in gene expression; the toggle-switch, in which two repressors inhibit each other's production to achieve a bistable system; the repressilator, in which three repressors are connected in a ring topology to produce repeated oscillation. "Rational" and "directed evolution" are currently used Genetic-circuit design tools. Someday we may be able to program cell behavior as easily as we program computers.

Construction and identification of synchronous nitric oxide-amplified gene circuits controlled by radiation / 中国肺癌杂志

<p><b>BACKGROUND</b>It has been proven that the positive feedback gene circuits can increase the expression level of interested genes, and the synchronization of genetic circuits can further enhance the efficacy of gene therapy. In order to obtain an enhanced and prolonged gene expression in target cells, a radiation controlled positive feedback genetic circuit is constructed via linking the c-fos promoter with the inducible nitric oxide synthase (iNOS) cDNA, which can be synchronized by nitric oxide (NO) intercellular messenger. Ultimately, the efficacy of radiogenetic therapy for cancer will be improved.</p><p><b>METHODS</b>Using the gene recombination techniques, the vector pfos-iNOS/green fluorescent protein (GFP) was generated by cloning the radiation-responsive c-fos promoter into the plasmid vector pIRES2-EGFP to replace the primary CMV promoter, and then inserting human iNOS cDNA downstream of c-fos promoter in the vector pIRES2-EGFP. The constructed plasmids were then downloaded into A549 cells with lipofectamine. With exposure of various doses of ionizing radiation, outputs of GFP and iNOS in the treated cells were observed and analyzed.</p><p><b>RESULTS</b>The interested plasmid was successfully constructed, proved by restriction enzyme digestion analysis. The outputs of GFP and iNOS in the transfected cells were markedly increased compared with the control cells after radiation, the peak level was seen in 16 hours after radiation.</p><p><b>CONCLUSIONS</b>A positive feedback genetic circuit is successfully developed, composed by c-fos promoter and iNOS cDNA, which can be synchronized by secreting the intercellular messenger NO. This genetic circuit will be utilized in further study.</p>

Study on enhancing sensitivity of SPC-A1 cells to chemotherapy by Livin isoform-specific gene silencing / 中国肺癌杂志

<p><b>BACKGROUND</b>As a new member of inhibitor of apoptosis protein(IAP) family,Livin,especially Livin α,is known to be involved in occurrence and development of lung cancer.Livin is an important mechanism of chemotherapy resistance of lung cancer cell.The aim of this study is to set up Livin isoform(α & β)-specific gene silencing system in SPC-A1 cells by gene transfection and RNA interference(RNAi),and to explore the different functions and value of the isoforms in enhancing chemosensitivity of SPC-A1 cells.</p><p><b>METHODS</b>Livinα+β,Livinα and Livinβ specific siRNA were expressed stably in SPC-A1 cells,respectively.MTT was performed to study sensitivity of the cells to chemotherapy drugs.In vivo experiment was performed to test sensitivity of mouse bearing tumor to cisplatin after gene silencing of Livin.</p><p><b>RESULTS</b>After silencing of Livinα+β,Livinα and Livinβ genes,sensitivity of SPC-A1 cells to many chemotherapy drugs(including cisplatin,carboplatin,cyclophosphamide and adriblastine) was markedly increased(P < 0.05).Among them,gene silencing of Livinα+β showed the strongest enhancement effect on chemosensitivity of SPC-A1 cells(P < 0.01).Animal experiment showed that tumor inhibition rate of pSilencer-Livinα+β,pSilencer-Livinα and pSilencer-Livinβ groups was 146.1%,130.7% and 110.5%,respectively.</p><p><b>CONCLUSIONS</b>The results suggest that Livin isoform,especially Livinα+β is hopeful to be a molecular target for increasing sensitivity of lung cancer cell to chemotherapy.Gene silencing may be a new means of gene therapy for non-small cell lung cancer.</p>

The expression of human antigen R and vascular endothelial growth factor-C and their significance in non-small cell lung cancer / 中国肺癌杂志

<p><b>BACKGROUND</b>Vascular endothelial growth factor-C(VEGF-C) plays a critical role in tumor-induced lymphangiogenesis and contributes to lymph node metastasis.Human antigen R(HuR) is one of the firstly identified RNA-binding proteins.It can increase the stability of a variety of growth factors and cytokines and upregulate protein expression.The aim of this study is to investigate the expression of HuR and VEGF-C protein in non-small cell lung cancer(NSCLC),and explore the relationship between the expression of HuR and VEGF-C and clinicopathological factors.</p><p><b>METHODS</b>HuR and VEGF-C protein levels were detected in 81 NSCLC tissues and 15 control benign pulmonary lesion tissues by immunohistochemistry method(SP method).</p><p><b>RESULTS</b>In NSCLC tissues,positive rate of cytoplasmic HuR,nuclear HuR and VEGF-C was 45.7%(37/81),82.7%(67/81) and 70.4%(57/81),respectively.There was a significant difference in positive expression of HuR and VEGF-C between NSCLC and benign pulmonary lesion tissues(P < 0.05).The expression of cytoplasmic HuR was closely related to pTNM stages,differentiation degree and lymph node metastasis(P < 0.05),but not correlated with sex,age and histological classification(P > 0.05).Furthermore,cytoplasmic immunoreactivity for HuR protein(P < 0.05) but not nuclear HuR expression(P > 0.05) was associated with high VEGF-C expression.</p><p><b>CONCLUSIONS</b>Cytoplasmic HuR and VEGF-C are overexpressed in NSCLC,and are related to tumor development.HuR may mediate the modulation of VEGF-C gene expression in NSCLC.</p>

Inhibitory effects of shRNA targeting layilin on adhesion and invasion behavior of human lung adenocarcinoma A549 cells induced by hyaluronan in vitro / 中国肺癌杂志

<p><b>BACKGROUND</b>It has been proved that hyaluronan was involved in adhesion and invasion behavior of varied tumor cells. Layilin is a receptor of hyaluronan found recently and has close relationship with cytoskeleton. The aim of this study is to investigate the effect of short hairpin RNA (shRNA) targeting layilin on adhesion and invasion behavior of human lung adenocarcinoma cell line A549 induced by hyaluronan in vitro.</p><p><b>METHODS</b>RNA interference plasmid that included U6 promoter and could express shRNA targeting layilin was designed, constructed, and transfected into A549 cell line. Layilin expression was examined by RT-PCR, Western blot and immunofluorescence. Adhesive and invasive ability was examined by plate adhesion model and Boyden chamber model.</p><p><b>RESULTS</b>After plasmid transfected, layilin expression in A549 cells obviously decreased (P < 0.01), and the numbers of adhesive A549 cells on plate and A549 cells permeating septum of Boyden chamber induced by hyaluronan also significantly decreased (P < 0.01).</p><p><b>CONCLUSIONS</b>The shRNA targeting layilin can efficiently inhibit the adhesive and invasive ability of A549 cells induced by hyaluronan in vitro.</p>

Study on relationship between VEGF-C and lymphangiogenesis and lymph node metastasis in non-small cell lung cancer / 中国肺癌杂志

<p><b>BACKGROUND</b>Increasing evidences have shown that vascular endothelial growth factor C (VEGF-C) is involved in the tumor lymphangiogenesis via combining with its ligand, vascular endothelial growth factor receptor-3 (VEGFR-3), which is the most important factor contributing to lymph node metastasis. Futhermore, lymph node metastasis is the most familiar metastatic pathway in non-small cell lung cancer (NSCLC). Based on this knowledge, the present study aims to explore the relationship between VEGF-C and lymphangiogenesis and lymph node metastasis in NSCLC with the use of podoplanin, a novel lymphatic vessel endothelium marker to detect lymphatic vessel.</p><p><b>METHODS</b>The expression of VEGF-C and podoplanin were detected in 66 paraffin sections of NSCLC and 8 inflammatory pseudotumor tissues by immunohistochemistry (SP). Assessments of podoplanin+ lymphatic vessel density (LVD) and positive rates of VEGF-C were performed and their relationship was analysed.</p><p><b>RESULTS</b>The positive rate of VEGF-C in the inflammatory pseudotumor group (12.5%) was significantly lower than that in the NSCLC group (75.8%) (P < 0.01), the positive rate of VEGF-C in the positive lymph node group was significantly higher than that in the negative lymph node metastasis group (86.5% vs 62.1%, P < 0.05) and there was no difference between the well differentiated group and the poor differentiated group (76.3% vs 75.0%, P > 0.05). LVD in the positive VEGF-C group was significantly higher than that in the negative VEGF-C group (21.3±6.0 vs 17.7±5.1, P < 0.05), LVD in the inflammatory pseudotumor group (10.9±4.9) was distinctly lower than that in the NSCLC group (20.4±5.9) (P < 0.01) and compared with the negative lymph node metastasis group (18.5± 5.5), LVD (21.9±5.9) in the positive lymph node metastasis group increased significantly (P < 0.05).</p><p><b>CONCLUSIONS</b>Lymphangiogenesis is a significant factor for tumor lymphatic metastasis, VEGF-C may mediate lymphatic metastasis of NSCLC by the way of inducing lymphangiogenesis.</p>

Construction of a liver targeting gene transfer vector using hepatitis B virus envelope particles and assessment of its function / 中国肿瘤生物治疗杂志

Objective: To construct a liver targeting gene transfer vector using hepatitis B virus envelope particles. Methods: Hepatitis B viruses were obtained from the supernatant of HepG 2.2.15 cells by a PEG8000 system and were inactivated by ?-propiolactone to prepare hepatitis B virus envelope. The hepatitis B virus envelope was used to pack 5.3 kb pIRES_2-EGFP to assess their packing ability. Subsequently, the products were studied with ELISA, PCR, SDS-PAGE, and electron microscopy. Finally, the product was used to transfect HepG2 cells and the green fluorescent protein (GFP) expression was observed under a fluorescent microscope. The rate of GFP positive cells was determined by flow cytometer.Results: The acquired hepatitis B virus envelope retained the surface protein HBsAg+pre S_1+pre S_2, but with no virus DNA. The prepared envelope had high packing ability for GFP and the packed GFP had a high transfection rate in HepG2 cell. Conclusion: Hepatitis B virus envelope has been successfully obtained from the supernatant of HepG 2.2.15 cells with a PEG8000 system and ?-propiolactone.

A randomized, prospective, multi-centre clinical trial of NP regimen (vinorelbine+cisplatin) plus Gensing Rg3 in the treatment of advanced non-small cell lung cancer patients / 中国肺癌杂志

<p><b>BACKGROUND</b>Gensing Rg3 is an active component from ginseng. The aim of this study is to observe the clinical anticancer effect of Rg3 in combination with chemotherapy regimen NP (vinorelbine+cisplatin) in advanced non-small cell lung cancer (NSCLC).</p><p><b>METHODS</b>Stage III-IV NSCLC patients confirmed by pathology or cytology all received vinorelbine plus cisplatin for at least two cycles, and were randomized into two groups: patients in arm A also received placebo twice a day, while patients in arm B received two tablets of Rg3 twice a day for at least two months. The endpoints of the study were the efficacy, survival and tolerance of patients.</p><p><b>RESULTS</b>From July 2000 to May 2002, 115 patients were enrolled into the trial. The patients' characteristics were well balanced in the two groups. Sex of patients: male, 79; female 36. Types of pathology: adenocarcinoma, 71; squamous cell carcinoma, 29; adenosquamous carcinoma, 8; others, 7. TNM stage: stage III, 45; stage IV, 70. Prior chemotherapy: with, 17; without, 98. Prior radiotherapy: with, 15; without, 100. Prior surgical treatment: with, 23; without, 92. Nine patients discontinued from the trial due to severe adverse effects (5) and other reasons (4), so there were 106 patients evaluable for clinical efficacy. The response rate was 14.5% (8/55) in arm A, and 33.3% (17/51) in arm B (P=0.011). The survival time in arm A was 9.7 months (mean) and 8.0 months (median), and 15.3 months (mean) and 10.0 months (median) in arm B (P=0.0088).</p><p><b>CONCLUSIONS</b>Preliminary results show improvements in response rate and survival time (median and mean) in Rg3 arm compared with placebo arm. It is worthy to confirm the results in further clinical trials.</p>

Correlation between expression of lung resistance-related protein in non-small cell lung cancer and that in peripheral blood lymphocytes / 第三军医大学学报

Objective To explore the correlation between the expression of lung resistance-related protein (LRP) in non-small cell lung cancer (NSCLC) and that in peripheral blood lymphocytes (PBL). Methods The S-P immunohistochemistry was used to detect the expression of LRP in specimens and PBL of 49 patients with NSCLC. Para-carcinoma tissues and PBL of 10 normal people were used as controls. Results The positive expression rate of LRP was 78.2%, 46.4% and 66.8% in carcinoma tissues, para-carcinoma tissues and PBL, respectively. The expression rate of LRP in carcinoma tissues was higher than that in para-carcinoma tissues (P

Neoadjuvant chemotherapy,adjuvant chemotherapy and radiotherapy or their combinations for nasopharyngeal cancer:a Meta-analysis on randomized controlled trials / 第三军医大学学报

Objective To compare the beneficial efficiency of chemotherapy combined with radiotherapy or radiotherapy alone,for locally advanced nasopharyngeal carcinoma.Methods A systematic review of randomized controlled trials(RCT) on this subject was performed.Mortality rate(MR),event-free survival(EFS),loco-regional control rate(LRR) and distant metastasis rate(DMR) were analyzed.Meta-analysis was carried out to compare the efficiency of adding chemotherapy to radiotherapy or radiotherapy alone.Results Totally 16 trials involving 3 768 patients were included.Our results showed that:① Patients receiving concomitant chemotherapy had better 2-,3-and 5-year MR,EFS,LRR and DMR compared with those receiving radiotherapy alone;② Neoadjuvant chemotherapy improved LRR and DMR in patients with nasopharyngeal carcinoma when compared with radiotherapy alone;③ There was no significant difference between adjuvant chemotherapy and radiotherapy alone in MR,EFS,LRR and DMR.Conclusion Adding chemotherapy to radiotherapy improves survival rate and EFS of patients with nasopharyngeal carcinoma,reduces LRR and DMR of these patients when compared with radiotherapy alone.We believe that adding chemotherapy to radiotherapy is a very effective therapy for patients with locally advanced nasopharyngeal carcinoma.

Inhibitory effects of shRNA targeting CD44v3 on invasion behavior of human colon cancer cell SW480 in vitro induced by hyaluronan / 中国癌症杂志

Purpose:To investigate the effect of short hairpin RNA (shRNA) targeting CD44v3 on adhesion and invasion behavior of human colon cancer cell line SW480 induced by hyaluronan in vitro. Methods:RNA interference plasmid including U6 promoter and expressing shRNA of CD44v3 was designed, constructed, and transfected into SW480 cell line. CD44v3 expression was examined by reverse transcription polymerase chain reaction (RT-PCR) and Western blot. Adhesive and invasive ability were examined by plate adhesion model and Boyden chamber model.Results:After plasmid transfection,CD44v3 expression in SW480 cell and the number of SW480 cell induced by hyaluronan adhesiving on plate or permeating septum of Boyden chamber decreased significantly (P

Apoptosis of lung adenocarcinoma SPC-A1 cells induced by Livin isoform-specific gene silencing / 中国癌症杂志

Purpose:To explore different functions and values of Livin ? and Livin ?-specific silencing in inducing SPC-A1 cell apoptosis, respectively. Methods:First, Livin ?+?, Livin?and Livin ?-specific siRNA were steadily expressed in SPC-A1 respectively. Next, after isoform-specific gene silencing, biological changes with regard to cell morphology and proliferation of lung cancer cell were observed. Then, TUNEL and FCAS were performed to test the rate of apoptosis. Lastly, statistical analysis was performed to explore the different functions and values of isoform-specific gene silencing in inducing lung cancer cell apoptosis.Results:After gene silencing of Livin ?+?, Livin ? and Livin ?,there was an evident decrease in colony forming ability of SPC-A1 compared with the control(P

Whether the hepatitis B virus envelope as a gene transfer vector has targeting ability for liver cancer / 中国癌症杂志

Background and purpose:The reason for hepatitis B virus (HBV) with hepatocyte specificity is PreS1 enchased on the hepatitis B virus envelope (HBVE). So HBVE may have a potential application in liver targeting gene transfer. In this study, we investigated whether HBVE has the ability to target liver cancer cells. Methods:HBVE was obtained from the supernatant of Hep G 2.2.15 cells through PEG8000 system and ?-propiolactone method. The pIRS2-EGFP was packed with HBVE and resulted in the product HBVE-GFP. HBVE-GFP was transfected into HepG2, A549, HeLa and FB cells. The green fluorescent protein (GFP) expression was observed under a fluorescent microscope. The rate of GFP positive cells was determined by flow cytometer.Results:The GFP could be observed in the four groups, but the HepG2 group had a higher fluorescent intensity than the other 3 groups. The transfected rate of HepG2 group was (71.35?0.03)% , much higher than other groups(P

EFFECTS OF MRF4 TRANSFECTION ON DIFFERENTIATION AND EXPRESSION OF MYOGENIC REGULATORY FACTORS IN CULTERED HUMAN RHABDOMYOSARCOMA RD CELLS / 解放军医学杂志

To study the effects of MRF4 transfection on differentiation and expression of myogenic regulatory factors of human rhabdomyosarcoma RD cells, the plasmid-MRF4 cDNA was transfected into cultured rhabdomyosarcoma RD cells with lipofectin method. The myogenic regulatory factors MRF4 and MyoD mRNA were measured with in situ hybridization and the expressions of myosin heavy chain(MHC) and a-actin in the cells were assayed with immunocytochemical method. The cell growth and morphology were observed at the same time. It was found that the morphology of differentiation increased and the growth was suppressed in RD cells after transfection. The expression of MHC and a-actin were significantly increased in RD cells after transfection, while the expressions of MRF4 and MyoD mRNA were up-regulated. It is suggested that transfection of MRF4 can induce differentiation of RD cells and up-regulate the expression of MyoD.

Expression, purification and identification of recombinant signal peptide of human ?-defensin 2 in A549 cells / 第三军医大学学报

Objective To construct high expression system for obtaining human ?-defensin 2 (h?D2) in human lung cancer cell line A549. Methods A recombinant retrovirus expression vector pLNCX2-CEA (signal peptide)-h?D2 (mature peptide) was constructed, and then the retrovirus vector was transfected into PT67 cells by DOTAP. After screening and amplification of single clones by G418, the virus was used to infect A549 cells. A549 cells were also screened by G418, and the cell clones resistant to G418 were obtained. Expression of h?D2 was detected by Western blot analysis. Results A recombinant retrovirus expression vector pLNCX2-CEA-h?D2 was constructed successfully. h?D2 with high level of expression was obtained in A549 cells transfected with the expression vector. Conclusion Human lung cancer cell line A549 can be used for high expression of h?D2 with gene engineering technology.

Changes of reactive oxygen species and mitochondrial membrane potential in mitochondrial DNA-depleted SK-Hep1 cells / 第三军医大学学报

Objective To observe the differences of the reactive oxygen species(ROS) and mitochondrial membrane potential(??m) in mitochondrial DNA-depleted SK-Hep1 and its parent cells.Methods ?0SK-Hep1 cell line was generated by treating SK-Hep1 with ethidium bromide.Flow cytometry(FCM) and laser scanning confocal microscopy were used to detect ROS and ??m.Results Fluorescence intensity of ROS in ?0SK-Hep1 cells was significantly higher than that of the parent cells(35.5 vs 15.6),and the ??m was lower in ?0SK-Hep1 cells(55.0 vs 65.9).Conclusion ?0SK-Hep1 cells showed elevated ROS and lower ??m.These changes might contribute to the variation of phenotypes.