MiR-3653 blocks autophagy to inhibit epithelial-mesenchymal transition in breast cancer cells by targeting the autophagy-regulatory genes ATG12 and AMBRA1 / 中华医学杂志(英文版)

BACKGROUND@#Metastasis is the main cause of tumor-associated death and mainly responsible for treatment failure of breast cancer. Autophagy accelerates tumor metastasis. In our work, we aimed to investigate the possibility of microRNAs (miRNAs) which participate in the regulation of autophagy to inhibit tumor metastasis.@*METHODS@#MiRNA array and comprehensive analysis were performed to identify miRNAs which participated in the regulation of autophagy to inhibit tumor metastasis. The expression levels of miR-3653 in breast cancer tissues and cells were detected by quantitative real-time polymerase chain reaction. In vivo and in vitro assays were conducted to determine the function of miR-3653. The target genes of miR-3653 were detected by a dual luciferase reporter activity assay and Western blot. The relationship between miR-3653 and epithelial-mesenchymal transition (EMT) was assessed by Western blot. Student's t -test was used to analyze the difference between any two groups, and the difference among multiple groups was analyzed with one-way analysis of variance and a Bonferroni post hoc test.@*RESULTS@#miR-3653 was downregulated in breast cancer cells with high metastatic ability, and high expression of miR-3653 blocked autophagic flux in breast cancer cells. Clinically, low expression of miR-3653 in breast cancer tissues (0.054 ± 0.013 vs . 0.131 ± 0.028, t  = 2.475, P  = 0.014) was positively correlated with lymph node metastasis (0.015 ± 0.004 vs . 0.078 ± 0.020, t  = 2.319, P  = 0.023) and poor prognosis ( P  < 0.001). miR-3653 ameliorated the malignant phenotypes of breast cancer cells, including proliferation, migration (MDA-MB-231: 0.353 ± 0.013 vs . 1.000 ± 0.038, t  = 16.290, P  < 0.001; MDA-MB-468: 0.200 ± 0.014 vs . 1.000 ± 0.043, t  = 17.530, P  < 0.001), invasion (MDA-MB-231: 0.723 ± 0.056 vs . 1.000 ± 0.035, t  = 4.223, P  = 0.013; MDA-MB-468: 0.222 ± 0.016 vs . 1.000 ± 0.019, t  = 31.050, P  < 0.001), and colony formation (MDA-MB-231: 0.472 ± 0.022 vs . 1.000 ± 0.022, t  = 16.620, P  < 0.001; MDA-MB-468: 0.650 ± 0.040 vs . 1.000 ± 0.098, t  = 3.297, P  = 0.030). The autophagy-associated genes autophagy-related gene 12 ( ATG12 ) and activating molecule in beclin 1-regulated autophagy protein 1 ( AMBRA1 ) are target genes of miR-3653. Further studies showed that miR-3653 inhibited EMT by targeting ATG12 and AMBRA1 .@*CONCLUSIONS@#Our findings suggested that miR-3653 inhibits the autophagy process by targeting ATG12 and AMBRA1 , thereby inhibiting EMT, and provided a new idea and target for the metastasis of breast cancer.

Association study of genetic variations in SLCO1B3 gene with prognosis in breast cancer patients treated with neoadjuvant chemotherapy of TA regimen / 中华肿瘤杂志

Objective@#To assess the association of single nucleotide polymorphisms (SNPs) in SLCO1B3 gene with prognosis of breast cancer (BC) patients treated with neoadjuvant chemotherapy of TA regimen (taxane and antharcycline drugs).@*Methods@#439 female BC patients were recruited and treated with neoadjuvant chemotherapy of TA regimen. A blood sample (2 ml) of peripheral blood was collected from each patient before chemotherapy. Tagging SNPs (tag-SNPs) were selected. We investigated the association of tag-SNPs with prognosis, by Sequenom Mass ARRAY system platform, characterizing tag-SNPs. The hazard ratio (HR) and 95% confidence interval (CI) for progression or death were calculated by multivariable-adjusted Cox regression model.@*Results@#Seven tag-SNPs (rs11045689, rs200104106, rs3764006, rs3834935, rs4149117, rs7305323 and rs73241801) were selected for study. Compared with individuals carrying the rs11045689 GG genotype, individuals carrying rs11045689 AA genotype performed worse PFS and OS, with the HR (95% CI) for progression being 1.39 (1.11~1.75) and the HR (95% CI) for death being 1.38 (1.04~1.83). Compared with individuals carrying the rs73241801 CC genotype, individuals carrying rs73241801 TT genotype performed better OS (P=0.041), with the HR (95% CI) for death being 0.65 (0.44~0.94). The number of risk allele was significantly associated with PFS (P=0.012) and OS (P=0.017) of BC patients by accumulation analysis. Compared with individuals carrying one or less than one risk allele, individuals carrying four risk alleles performed worse PFS and OS, with the HR (95% CI) for progression being 1.37 (1.09~1.72) and the HR (95% CI) for death being 1.36 (1.02~1.81).@*Conclusion@#The variations of rs11045689 and rs73241801 in SLCO1B3 gene were significantly associated with prognosis of BC patients treated with neoadjuvant chemotherapy of TA regimen, which might serve as biomarkers for predicting prognosis of BC patients treated with neoadjuvant chemotherapy.

The significance of postoperative radiotherapy in uterine sarcoma / 中华放射肿瘤学杂志

Objective To evaluated the role of postoperative radiotherapy (PR) after surgery in patients with uterine sarcoma,and analyzed the prognostic factors.Methods A total of 182 patients with uterine sarcoma were included between June 1994 and October 2014.Radiotherapy dose were 30-50 Gy/10-25 fractions/5 fractions/week.The LRFFS and OS were calculated with Kaplan-Meier method,and difference was analyzed with log-rank method.Cox regression analyses were used to determine prognosticators.Results There were 114 patients which survived more than 5-years in this whole group,including PR 24 cases and no PR 90 cases.The 5-year LRRFS and OS were 62.1％ and 56.2％,respectively.The 5-year LRRFS were 78.0％ and 55.3％ on PR and no PR (P=0.013);with OS 64.1％ and 51.7％ on PR and no PR (P=0.070).A multivariate analysis showed that pathological types,histological grade and clinical stage were associated with LRRFS and OS (P=0.032,0.008,0.000 and 0.046,0.000,0.000).PR was significant influencing factor for OS (P=0.013).Conclusions Uterine sarcoma patients treated with PR after surgery had an improved LRRFS compared to those treated with surgery,especially those with leiomyosarcoma.The role of PR personalized radiation for uterine sarcoma still needs to be further discussed.

Impact of stages of axillary lymph nodes and pathological response to neoadjuvant chemotherapy on the survival of breast cancer patients / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To analyze the relevance between lymph node status and pathological response after neoadjuvant chemotherapy and survival in breast cancer patients.</p><p><b>METHODS</b>The clinicopathological data of 653 needle biopsy proved breast cancer patients, who underwent neoadjuvant chemotherapy and surgery in our hospital from July 1998 to April 2012, were retrospective analyzed.</p><p><b>RESULTS</b>The median follow up time was 59.3 months. The 653 cases were classified into ypN0 (242 cases), ypN1 (182 cases), ypN2 (135 cases), and ypN3 (94 cases) stages, and the 5-year overall survival rates in the four groups were 93.4%, 93.4%, 87.4%, and 83.0%, respectively. The Log rank test showed a significant difference in the overall survival rates between the ypN0, ypN1, ypN2 stages and ypN3 stage (P=0.046). No significant differences were observed between the disease free survival (DFS) rates in the four groups (P>0.05). Multivariate analysis indicated that the postoperative pathological response of metastatic lymph nodes was a major prognostic factor affecting the overall survival and disease-free survival (RR=1.051, P=0.007; RR=1.028, P=0.028).</p><p><b>CONCLUSION</b>The stage and pathological response of axillary lymph nodes after neoadjuvant chemotherapy are effective indicators for predicting the OS and DFS in breast cancer patients.</p>

Diagnostic value of X-ray stereotactic vacuum-assisted biopsy for breast micro-calcification / 肿瘤研究与临床

Objective To compare the accuracy and clinical effect between X-ray stereotactic vacuum-assisted biopsy (SVAB) and metal wire guided lumpectony.Methods From January 2010 to June 2014,681 cases of breast micro-calcification biopsy were performed.Among them,78 cases were performed with SVAB and 603 eases were performed with the method of stereotaetic metal wire guided lumpectomy.All cases were non-palpable breast lesions (NPBLs) and breast imaging-reporting and data system (BI-RADS) assessment categories 4.The diagnostic accuracy and clinical effect were compared.Results The sensitivity of both methods was 100 ％ with no misdiagnosis.The underestimation rate of SVAB was 12.5 ％.Compared with the method of metal wire guided lumpectomy,SVAB had many advantages,such as easy to use,quickly performed,low rate of local deformation and lower rate of operative complications.77.5 ％ patients benefited from SVAB by avoiding open surgery of benign disease.Conclusions SVAB is an accurate,safe and convenient method of biopsy.It can be recommended as the preferred method of micro-calcification (BI-RADS 4).Additional operation should be performed on patients with the pathological diagnosis of middle and high grade of dysplasia and any kind of carcinoma.

Genomics of breast cancer / 中国肿瘤临床

Breast cancer is caused by somatic mutation. As such, somatic mutation in breast cancer should be described to eluci-date the underlying mechanism. Next-generation sequencing has provided new insights into the genomics of breast cancer. New genes were identified and exhibited a relationship with breast cancer. Although these genes mutated at a low frequency, such genes in different cases could be categorized into specific pathways. Mutational signatures could be found in some cases, but such signatures were gener-ally not related to environmental exposure. Studies on intra-tumoral heterogeneity have revealed the ubiquitous presence of sub-clones in breast cancer;however, a major clone is also observed, accounting for>50%of tumor cells. Current advancements show that breast cancer genomics has been integrated into personalized medicine. Furthermore, a genome-informed and personalized molecular sub-typ-ing and treatment of breast cancer can be developed in the future.

Effect of Cancer Target Drug LHRH-PE40 on Elasticity of HeLa Cells / 分析化学

The quantitative analysis of biomechanical profiles at the single-cell level can provide additional information. It is usually not available in traditional cell biology approaches, but may be crucial to assess and understand tumor prognosis and response to chemotherapy. In this study, the on-line changes of cell elastic properties after the addition of cancer target drug LHRH-PE40 were monitored by atomic force microscopy ( AFM) on living HeLa cell surface under physiological condition. The results from AFM based force spectroscopy showed that LHRH-PE40 induced a distinct increase of the cell surface elasticity of HeLa cells. The fluorescence images implied that the target drug LHRH-PE40 would affect the re-organization of cell actions, which led to the increase of the elasticity of HeLa cells.

Tumor radiosensitivity predicted by 18F-FLT PET imaging / 中华核医学与分子影像杂志

Objective To evaluate if 18F-FLT PET imaging could be used as a new clinical method to predict tumor radiosensitivity.Methods MDA-MB-231 and LN229 cells were irradiated with doses of 0,8 and 16 Gy of 6 MV photon energy,then soft agar assay and cellular uptake of 18F-FLT were performed on the 2 cell lines.The t test and one-way analysis of variance were used for the two groups and data before and after irradiation.The MDA-MB-231 and LN229 tumor xenografts were prepared by injecting the tumor cells into the right limbs of female BALB/c nu/nu mice.Once tumors reached a diameter of 10 mm,the two types of mice were divided randomly into 3 groups (20 mice per group) according to the irradiation doses (0,8 and 16 Gy).After irradiation,18F-FLT PET imaging and immunohistochemical staining were conducted.Then correlations between 18F-FLT SUVtumor/SUVmuscle ratio (T/M ratio) and TK1 labeling index percentages (LITK1) were tested using linear correlation analysis.Results The survival fraction of MDA-MB-231 and LN229 cells after irradiated with 8 Gy were (59.73 ± 4.3) ％ and (93.41 ± 3.75) ％,respectively (t =-13.20,P ＜ 0.001).When the dose increased to 16 Gy,the survival fraction decreased to (43.57 ±4.06) ％ and (81.77 ± 4.42) ％,respectively(t =-14.24,P ＜ 0.001).In MDA-MB-231 cells,the cellular uptake of 18F-FLT after irradiation with 8 Gy declined rapidly to (18.32 ± 1.38) kBq/105 cells ((128.22 ± 8.24) kBq/105 cells with the dose of 0 Gy,F =266.41,P ＜ 0.01),and maintained this low level till 72 h.For the LN229 cells,the cellular uptake decreased to (9.87 ± 1.30) kBq/105 cells after 8 Gy irradiation ((134.88 ± 6.59) kBq/105 cells with the dose of 0 Gy,F =346.06,P ＜ 0.01),then increased gradually to (127.17 ± 9.08) kBq/105 cells at 72 h (F =346.06,P ＞ 0.05).The dynamic changes of 18F-FLT cellular uptake in the two cells had the same pattern after being treated with 16 Gy irradiation.In the 18F-FLT PET image of MDA-MB-231 tumor mice after 8 Gy radiotherapy,the T/M ratio decreased to 0.78 ± 0.39 at the first day,but it was 2.84 ± 0.29 before radiotherapy (F =39.78,P ＜0.01).Then the ratio increased slowly,and it was still lower than the baseline at 7 d after radiation (F =39.78,P ＜0.01).The same pattern could be seen in the group of 16 Gy irradiation.In LN229 tumor mice treatment with 8 Gy irradiation,the T/M ratio increased to 2.41 ±0.47 at the first day,and it was 1.58 ±0.29 before radiotherapy (F =34.01,P ＜ 0.05).The ratio decreased steadily to 0.66 ± 0.32 (F =34.01,P＜0.05) at 7 d after radiotherapy.However,in the treatment group with 16 Gy,the T/M ratio decreased gradually and reached 0.44 ± 0.22 at 7 d (F =41.85,P ＜ 0.01).A correlation was found between 18F-FLT T/M ratio and LITK1 (8 Gy:r=0.67,0.73; 16 Gy:r=0.73,0.69; all P＜0.01) in both tumor models.Conclusion 18F-FLT PET imaging may be used as a new assay to predict tumor radiosensitivity,but further investigation is needed before clinical application.

The clinical characteristics and prognostic factors of aggressive fibromatosis in 142 patients / 中华放射肿瘤学杂志

Objective Aggressive fibromatosis is a rare kind of soft tissue tumor and was evaluated by few large studies. This study was to evaluate the clinical characteristics and identify the prognostic factors of this disease. Methods One hundred and forty-two patients with aggressive fibromatosis treated from January 1983 to August 2009 in Tianjin Medical University Cancer Hospital were retrospectively reviewed.The prognostic value of clinical and treatment factors was analyzed. Univariate analysis was performed with Log-rank test and Multivariate analysis was performed with Cox regression model. Results The follow-up rate is 93.7% and the median follow up time was 54 months (range,6 -208 months). Sixty-three patients had a minimum follow up time of 5 years and 6 patients had a minimum follow up time of 10 years. The male/female ratio was 1/1.84. The disease was most popular in women aged from 18 to 35 years old. The disease frequently occurred in the trunk (55.6%) and extremity (31.7%). All patients received surgery,and 46 received radiotherapy. The 5-year and 10-year local recurrence rates were 24. 4% and 31.1%,respectively. The 5-year and 10-year overall survival rates were both 99. 3%. Univariate analysis revealed that factors correlated with local recurrence were tumor size ( χ2 = 4. 37, P = 0. 037 ) and margin status (χ2 = 12. 36,P =0. 002). Multivariate analysis revealed that margin status was an independent risk factor (RR = 2. 219; χ2 = 9. 47,P = 0. 002) and radiotherapy was an independent protective factor ( RR = 0. 360;χ2 = 4. 95, P = 0. 026 ) for disease recurrence. When radiotherapy was delivered, the 10-year local recurrence rate decreased from 70. 1% to 20. 7% in patients with positive margin ( χ2 = 4. 22, P = 0. 040 )and decreased from 19.8% to 10.4% (χ2= 0.90, P= 0.344) in patients with negative margin.Conclusions Radical resection is the mainstay of treatment for aggressive fibromatosis. Postoperative radiotherapy can reduce the recurrent rate for patients with positive margin. In patients with negative margin,the role of radiotherapy should to be further evaluated in large clinical trials.

Prognostic factors of patients with locally recurrent rectal cancer after radical resection / 中华放射肿瘤学杂志

Objective To investigate the prognostic factors and the clinical outcome of locally recurrent rectal cancer after radical resection. Methods From April 2000 to April 2004, 105 patients with locally recurrent rectal cancer after radical resection were re-treated in Tianjin cancer hospital. Thirty-four patients were re-treated with surgery combined with adjuvant chemoradiotherapy (group 1), 35 with surgery alone (group 2), and 36 with chemoradiotherapy (group 3). The impact of 17 clinicopathological factors and treatment modalities on the survival was analyzed. Results The follow-up rate was 95. 2%. The median survival time was 23 months. The 1-, 3-and 5-year survival rates of patients with locally recurrent rectal cancer were 63% ,34% and 19%, respectively. The 1-, 3-and 5-year survival rates were 79%, 55% and 32% in group 1 ; 68%, 40% and 14% in group 2; and 64%, 36% and 11% in group 3; respectively (χ~2 =7. 96,P =0. 019). The univariate analysis showed that the degree of differentiation, depth of tumor invasion, number of metastatic lymph nodes, initial TNM stage, recurrent location, time to recurrence, and surgery combined with adjuvant therapy were significant prognostic factors, with the last 4 being the independent prognostic factors. Conclusions Surgery combined with chemoradiotherapy may improve the survival of patients with locally recurrent rectal cancer.

Core needle biopsy for the diagnosis of breast tumor / 中华普通外科杂志

Objective To evaluate the accuracy of core needle biopsy (CNB) in diagnosing breast masses and its coherence with immunohistochemical (IHC) examination results of estrogen receptor (ER), progesterone receptor (PR) and Her2 protein between pre-and post-chemotherapy in invasive breast cancer. Methods The results of 516 CNB cases from June, 2005 to April, 2008 were analyzed retrospectively. The pathological examination was performed by two pathologists independently. Results 484 cases of malignant tumor, carcinoma in situ and phyllodes tumor were found in this group with the sensitivity of 96.7%. Sixteen cases of false negative (3. 3% ) were demonstrated by surgical biopsy. The accurate rate of CNB was not influenced by the maximum diameter of masses ( P = O. 423 ). The agreement rate of IHC results of ER, PgR and Her2 between pre- and post-chemotherapy were 90. 3%, 76. 8% and 82.5%, respectively. Conclusion CNB is a useful diagnostic method with a satisfactory accuracy in any size of breast masses. Given the histological heterogeneity of invasive breast cancer and the influence of ehemotherapy, the coherence of prechemotherapy IHC for ER, PgR and Her2 is not optimal with that of post-chemotherapy.

Coadsorption of Adenine and Thymine on Au(111) Electrode / 分析化学

The coadsorption behavior of two complementary DNA bases,adenine(A) and thymine(T),was studied on Au(111) electrode by cyclic voltammetry(CV) and electrochemical scanning tunneling microscopy ( ECSTM) in aqueous solution. From the CVs,the coadsorption behavior of A and T was more closed to the adsorption behavior of A. In the potential range of physisorption,high- resolution ECSTM images of revealed that a new super-structure was formed by the hydrogen bonds between A and T. A molecular model of the super-structure was proposed based on the STM results and the possible hydrogen bonds between A and T.