Objective To investigate the effects and possible mechanisms of Mfn-2 gene overexpression on photodynamic therapy (PDT) sensitivity of T47D cells in human breast cancer.Methods pEGFP and pEGFP-Mfn-2 were transfected into human breast cancer T47D cells,and then the mRNA and protein expression of Mfn-2 gene in T47D cells were detected by real-time PCR (RT-PCR) and Western blotting in pEGFP group and EGFP-Mfn-2 group,respectively.After pEGFP and pEGFP-Mfn-2 were transfected into human breast cancer T47D cells for 48 hours,methyl thiazolil tetracolium (MTT) assay was used to measure the PDT sensitivity of T47D cells in human breast cancer in pEGFP group and pEGFP-Mfn-2 group.pEGFP + PDT group and pEGFP-Mfn-2 + PDT group were obtained by PDT irradiating pEGFP and pEGFP-Mfn-2 which were transfected into human breast cancer T47D cells.Cell apoptosis and mitochondrial membrane potential of T47D cells were assayed by flow cytometry in pEGFP + PDT group and pEGFP-Mfn-2 + PDT group.Laser scanning confocal fluorescence microscope was applied to observe the morphological ultrastructure of mitochondria in pEGFP group,pEGFP-Mfn-2 group,pEGFP + PDT group,and pEGFP-Mfn-2 + PDT group.Results RT-PCR showed that after transfecting T47D cells with pEGFP,the expression of Mfn-2 mRNA was 1.01 ±0.12.After transfecting T47D cells with pEGFP-Mfn-2,the expression of Mfn-2 mRNA was 1 067.00 ±41.72.There was statistical significance (t =67.541,P ＜ 0.001).Western blotting revealed that compared with the pEGFP group,the pEGFP-Mfn-2 transfection group had higher expression of Mfn-2 gene in T47D cells.MTT assay showed that pEGFP-Mfn-2 transfection significantly enhanced the PDT sensitivity of T47D cells in hunan breast cancer compared with the pEGFP + PDT group.When the concentration of methylene blue was 5.00 μmol/ml,the survival rate of the pEGFP +PDT group and pEGFP-Mfn-2 + PDT group were (59.96％ ± 1.21％) vs.(46.50％ ± 1.72％),with significant difference (t =34.403,P ＜ 0.001).Flow cytometry assay showed that the cell apoptosis rates in pEGFP-Mfn-2 + PDT group was markedly higher compared with the pEGFP + PDT group [(81.21 ± 2.13)％ vs.(68.82 ±2.64)％,P=0.024],with statistical significance.Also,the mitochondrial membrane potential was obviously lower in pEGFP-Mfn-2 +PDT group compared with the pEGFP +PDT group [(1.37 ±0.12)％ vs.(23.33 ± 1.86)％,P＜0.001],with statistical significance.Laser scanning confocal fluorescence microscope showed that cells in pEGFP group showed network structure,both pEGFP-Mfn-2 group and pEGFP + PDT group could cause mitochondrial fusion,and the pEGFP-Mfn-2 + PDT group could induce mitochondrial disintegration and lose its normal morphology completely.Conclusion Mfn-2 may enhance the PDT sensitivity of T47D cells in human breast cancer,which is possibly related with the normal morphology alteration of mitochondria and the inducement of mitochondrial apoptosis.

Objective To investigate the inhibitory effect of polyphyllin D on the proliferation of human chronic myelogenous leukemia (CML) cell line K562 cells and its mechanism.Methods Methyl thiazolyl thiazolium (MTT) assay was used to evaluate the inhibition effect of polyphyllin D on the proliferation of K562 cells.Flow cytometry was used to determine the effect of polyphyllin D on the apoptosis of K562 cells.The related proteins were analyzed by Western blotting.Results The MTT assay showed that polyphyllin D obviously inhibited the growth of K562 cells.The hemi-inhibitory concentration (IC50) of 24 hours for the K562 cells was (0.8 ± 0.1) μmol/L.Flow cytometric results demonstrated that polyphyllin D significantly induced apoptosis in a dose-dependent manner,and the apoptosis rates of 0,0.4,0.8,1.2 μmol/L polyphyllin D were (1.73 ± 0.33) ％,(14.47 ± 2.57) ％,(24.10 ± 3.52) ％,(31.51 ± 4.09) ％ after 24 hours,with a significant difference (F =58.231,P ＜ 0.001).Western blotting showed that polyphyllin D significantly down-regulated the expression of Bcl-2,and this was accompanied by up-regulation of Bax,decrease of the Bcl-2/Bax ratio,up-regulation of cytochrome c and activation of caspase-3.Otherwise,polyphyllin D obviously induced monocyte differentiation by increasing CD14 expression on the surface of K562 cells.Conclusion Polyphyllin D can significantly inhibit the proliferation of K562 cells,and the mechanism may be realized by inducing apoptosis and promoting monocyte differentiation.

Objective To investigate the expression and clinical significance of long non-coding RNA SBF2-AS1 (SBF2 antisense RNA 1) in non-small cell lung cancer (NSCLC).Methods Real-time quantitative polymerase chain reaction (qRT-PCR) was used to detect SBF2-AS1 expressions in 114 cases of NSCLC tissues and the adjacent normal tissues to analyze its relationship with clinic-pathological characteristics,diagnostic value and prognosis in NSCLC.Results SBF2-AS1 expression was significantly higher in the NSCLC (4.336 ±0.032) compared with the adjacent normal tissues (1.256 ± 0.021),with a significant difference (t =3.594,P =0.005).The expression of SBF2-AS1 was related with tumor size (x2 =13.072,P =0.001),lymphatic metastasis (x2 =6.896,P =0.009),TNM stage (x2 =8.566,P =0.003),smoking history (x2 =8.769,P =0.003) and infiltration degree (x2 =17.852,P =0.001),but was not related with age (x2 =0.141,P =0.707),sex (x2 =0.036,P =0.850) and pathological type (x2 =1.267,P =0.260).The area under the receiver operating characteristic (ROC) curve was 0.853 (95 ％ CI:0.755-0.879,P =0.004).The sensitivity and specificity was 52.4％ and 87.8％,respectively.The difference betwen low SBF2-AS1 expression and high SBF2-AS1 expression groups was statistically significant in overall survival time (42.3 months vs.25.2 months,x2 =4.753,P =0.013).Conclusion The expression of SBF2-AS1 is upregulated in NSCLC and may be proved useful as a biomarker and diagnostic target for the treatment of patients with NSCLC.

Objective To explore the effectiveness of epidermal growth factor receptor (EGFR)-tyrosine kinase inhibitors (TKIs) in non-small cell lung cancer (NSCLC) patients with EGFR double mutations containing rare mutations.Methods From March 2007 to January 2017,NSCLC patients with EGFR double mutations containing rare mutations confirmed by histopathology and EGFR mutation detections at Renmin Hospital of Wuhan University,Zhongnan Hospital of Wuhan University and Hubei Cancer Hospital were enrolled.According to the mutation types,patients were divided into double activating mutations group and activating mutations with insensitive mutations group.The effectiveness of TKIs in NSCLC patients with EGFR double mutations containing rare mutations and different mutation types was analysis.Results Among the 2 637 NSCLC patients underwent EGFR mutation detections,19 patients were confirmed as EGFR double mutations containing rare mutations.Fifteen patients received EGFR-TKIs therapy,the objective response rate (ORR),disease control rate (DCR) and median progression free survival (PFS) were 46.7％ (7/15),73.3％ (11/15) and 8.1 months,respectively.In patients with double activating mutations,two patients had partial response (PR),one patient had a stable disease (SD).In the activating mutations with insensitive mutations group,five patients had PR,three patients had SD,four patients had no effect.Conclusion Patients with EGFR double mutations containing rare mutations have a general response to EGFR-TKIs,and this result can provide a reference for the treatment of those patients.

Objective To evaluate the association between the decrease of the perioperative serum carcinoembryonic antigen (CEA) level during perioperative period and the prognosis in patients with colon cancer after a curative resection.Methods Retrospective analysis was conducted to evaluate the relationship between preoperative serum CEA level and different clinicopathologic features in 605 cases who underwent a curative resection for colon cancer from January 2006 to April 2011.According to the preoperative serum CEA level,the patients were divided into two groups:≤5 ng/ml and ＞5 ng/ml group.The critical value of the CEA decreasing rate in preoperative serum CEA ＞ 5 ng/ml group was calculated,and the relationship between this critical value and survival rate was then analyzed.Univariate and multivariate models were used to detect the risk factors of overall survival rate (OS) and disease free survival (DFS) in preoperative serum CEA ＞ 5 ng/ml patients.Results The preoperative serum CEA levels were significantly associated with lymphatic invasion (x2 =14.122,P＜0.001),T stages (x2 =40.153,P ＜0.001),N stages (x2 =22.721,P ＜0.001) and pathological stages (x2 =38.576,P ＜ 0.001),except for sex (x2 =0.453,P =0.501),age (x2 =0.195,P =0.659) and histological stages (x2 =6.135,P =0.112).The critical values of CEA decreasing rate for OS and DFS were 48.95％ and 50.81％ in preoperative serum CEA ＞5 ng/ml group respectively.There were significant differences of 5-year OS (31.37％ vs.76.63％,x2 =43.235,P ＜ 0.001) and 5-year DFS (27.69％ vs.72.10％,x2 =55.561,P ＜0.001) between patients after operation whose CEA decreasing rate were lower than critical value and those whose were higher.Univariate analysis showed that the decreasing rate of CEA was an influence factor for OS (x2 =43.235,P ＜ 0.001) and DFS (x2 =55.561,P ＜ 0.001) of preoperative serum CEA ＞ 5 ng/ml patients.The N stages and pathological stages were both related to OS (x2 =14.683,P＜0.001;x2 =12.295,P＜0.001) and DFS (x2 =16.212,P＜0.001;x2 =13.704,P＜0.001)respectively.Multivariate model showed that the decreasing rate of CEA level and N stages were both associated withOS (x2=18.885,P＜0.001;x2 =7.523,P＜0.001) and DFS (x2 =19.275,P＜0.001;x2 =6.997,P ＜ 0.001) of preoperative serum CEA ＞ 5 ng/ml patients.Conclusion A high decreasing rate of serum CEA level after operation in colon cancer patients who have high CEA levels before the curative resection can be a protective factor for prognosis,especially when the decreasing rate is higher than the critical value.

Objective To evaluate the expressions and prognostic significance of phosphorylation of protein kinase B1 (pAkt1) and human telomere reverse transcriptase (hTERT) in epithelial ovarian carcinoma.Methods The expressions of pAkt1 and hTERT were examined by immunohistochemical SP method in 72 cases of epithelial ovarian carcinoma and 10 normal endometrial tissues to analyze the correlation between pAkt1 and hTERT and their relationships with clinic-pathological features and prognosis.Results The positive expression rate of pAkt1 in epithelial ovarian carcinoma was 73.6％,and the positive expression rate of hTERT was 56.9％,compared with normal ovarian tissues,with statistical significance (x2 =20.814,P ＜ 0.001;x2 =11.3g9,P ＜ 0.001).The expression of pAkt1 was not associated with hTERT expression (r =0.075,P =0.532) in epithelial ovarian carcinoma.The expressions of pAkt1 and hTERT in epithelial ovarian carcinoma were correlated with tumor differentiation status (x2=6.475,P =0.011;x2 =1.370,P =0.001).The disease-free survival of patients with pAkt1 and hTERT positive coexpression was significantly shorter than others (8.585 months vs.11.227 months,x2 =4.361,P =0.037),but there was no significant difference in overall survival (11.107 months:11.695 months,x2 =0.394,P =0.530).Conclusion pAkt1 and hTERT are highly expressed in epithelial ovarian carcinoma,and high coexpression of pAkt1 and hTERT indicates poor prognosis in epithelial ovarian carcinoma.

Cancer stem cells have self-renewal and multilineage differentiation potentials,which play important roles in tumorigenesis,metastasis,recurrence and resistance to chemical therapy and radiotherapy,and so on.It is the hotspot of tumor research recently.Aptamer is a small molecular oligonucleotide with high targeting ability and affinity,which is a new type of nucleic acid in the diagnosis and treatment of cancer.Now more and more research is being done on the aptamer is applied in the diagnosis and treatment of cancer stem cells,which provides a new research tool for the diagnosis and treatment of cancer stem cells.

The abnormality of long interspersed nuclear element-1 (LINE-1) is closely related to the occurrence and development of tumor.Overexpression of LINE-1 can promote cell proliferation and affect the prognosis and metastasis of tumor.The retrotransposition and methylation status of LINE-1 have potential application value in clinical diagnosis,prognosis,disease monitoring and treatment.

Numerous studies support the importance of gut microbiota as environment factors in the development of cancer.The mechanisms that gut microbiota drive certain cancers include impacting immunity,modulating inflammation,inducing DNA damage,producing cancer promoting metabolites and regulating signaling pathway.Microbiome-based therapies provide potential prospects for cancer treatment.

Fenofibrate is a common lipid-lowing drug activating peroxisome proliferator-activated receptor alpha (PPRAα).Recent studies have found that fenofibrate possesses anticancer properties.Its specific mechanisms include inhibiting cell metabolism and formation of new tumor vessels,arresting cell circle,weakening cell motility,and inducing cell apoptosis.Those properties are partly independent of PPRAα.Due to its low side-effects,it's hopefully to be used as an anticancer adjuvant drug.