Effects of circ_0063865 silencing on the malignant phenotype of esophageal squamous cell carcinoma / 中华老年医学杂志

Objective:To investigate the expression of circ_0063865 in esophageal squamous cell carcinoma(ESCC)tissues and cells and its effect on the biological properties of the cells.Methods:The loop structure and stability of circ_0063865 were identified by Sanger sequencing, back-to-back primer validation and the ribonuclease R(Rnase R)tolerance assay.The expression of circ_0063865 was detected by RNA fluorescence in situ hybridization in an ESCC tissue microarray and its clinical relevance was analyzed.The expression levels of circ_0063865 in a normal esophageal epithelial cell line and ESCC cell lines were measured by real-time quantitative polymerase chain reaction(RT-qPCR). Cell counting Kit-8, the colony formation assay, the scratch assay, the transwell invasion assay and flow cytometry were used to detect the effects of circ_0063865 on cell proliferation, migration, invasion abilities and apoptosis, respectively.Results:The loop formation of circ_0063865 was verified by Sanger sequencing, back-to-back primer and Rnase R tolerance assays.The results of RNA fluorescence in situ hybridization showed that the mean fluorescence intensity of circ_0063865 expressed in ESCC tissues was significantly higher than in its paired paracancerous normal tissues( t=2.267, P<0.05). The expression of circ_0063865 was significantly associated with lymph node metastasis( χ2=4.356, P<0.05). The average overall survival time of patients with high circ_0063865 expression ESCC was lower than that of patients with low circ_0063865 expression ESCC.RT-qPCR results demonstrated that, compared with HEEC, circ_0063865 expression was elevated in ESCC cell lines( F=18.413, P<0.05). In addition, after circ_0063865 knockdown, the proliferation, migration and invasion abilities of KYSE-30 and KYSE-150 cells were significantly decreased, and the level of apoptosis was significantly increased(both P<0.05). Conclusions:The expression of circ_0063865 in ESCC is high, and changes in its expression are significantly correlated with lymph node metastasis.Additionally, circ_0063865 can promote the proliferation, migration and invasion of ESCC cells.

Effects of 5-Aza-dC combined with chemotherapy regimens on the apoptosis of lung adenocarcinoma cells / 中国综合临床

Objective:To investigate the effect of different chemotherapy drugs combined with DNA methylase inhibitor 5-Aza-2'-deoxycytidine (5-Aza-dC) on the apoptosis of lung adenocarcinoma cells.Methods:In the prospective randomized controlled study, lung adenocarcinoma A549 cells were treated with cisplatin plus paclitaxel (TP) or gemcitabine (GP) with or without 5-Aza-dC. According to different drug intervention methods, they were divided into control group, cisplatin combined with paclitaxel (TP) group, cisplatin combined with gemcitabine (GP) group, and 5-Aza-dC combined with TP group, 5-Aza-dC combined with GP group. CCK-8 assay was used to detect the proliferation of A549 cells. Transwell migration and invasion assay were used to detect the effect that each group of drugs on the migration and invasion ability of A549 cells. Quantitative Real-time Polymerase Chain Reaction was used to evaluate the effect of each treatment on the expression of apoptotic genes. One-way analysis of variance was used to compare the degree of cell proliferation in different drug treatment groups, and LSD- t method was used for pairwise comparison within groups. Results:The inhibition rates of lung adenocarcinoma cells in the TP regimen at different time points at 24, 48, and 72 h were as follows (20.00±4.23) %, (35.00±2.80) %, and (56.00±3.11) %. The inhibition rate of 5-Aza-dC combined with TP regimen on lung adenocarcinoma cells was significantly increased, at different time points of 24, 48 and 72 h, respectively (38.00±3.80) %, (50.00±3.25) %, (93.00±4.33) %. The inhibition rates of cells at different time points at 24, 48, and 72 h in the GP regimen were (33.00±5.10) %, (54.00±3.80) %, and (74.00±2.82) %, respectively; while 5-Aza-dC combined with GP regimen could significantly reduce the rate of cell growth, the inhibition rates of cells at 24, 48, and 72 h different time points were as follows (54.00±3.00) %, (67.00±5.30) %, and (95.00±1.13) %. The inhibitory effect of the same drug on lung adenocarcinoma cells increased with time (TP group: F=35.93, P<0.001; 5-Aza-dC combined with TP group: F=97.33, P<0.001; GP group: F =41.73, P<0.001; 5-Aza-dC combined with GP group: F=79.00, P<0.001), and at different time points, the differences were statistically different (all P<0.05). 5-Aza-dC combined with TP and GP chemotherapy regimens can inhibit the migration and invasion of lung adenocarcinoma cell A549, and the inhibitory effect is stronger than that of TP or GP regimens alone. The expression of Caspase 8 was significantly elevated ( t=5.87, P=0.004) in cells treated with 5-Aza-dC combined with GP when compared with GP regimen alone. The expression of Caspase 8 ( t=3.94, P=0.017), Caspase 6 ( t=5.81, P=0.004) and BBC3 (BCL-2 binding component 3) ( t=6.53, P=0.003) were increased when drugged with 5-Aza-dC combined TP regimen compared with TP regimen alone. Conclusion:5-Aza-dC might serve as a chemotherapeutic sensitizer to increase the sensitivity of lung adenocarcinoma cells.

Dawn of light: opportunities and challenges in small cell lung cancer immunotherapy / 中国综合临床

Small cell lung cancer accounts for about 15% of all lung cancers, and is a highly invasive neuroendocrine tumor. smoking is a major risk factor. SCLC grows rapidly, has a high metastasis rate and has a poor prognosis. For more than 30 years, the treatment of SCLC has progressed slowly, until the emergence of immunodrugs in recent years, which have achieved certain efficacy in a wide range of patients.

Application of combined body position fixation in radiation therapy of lung cancer / 中华放射医学与防护杂志

Objective To explore the CTV to PTV external expansion boundary and the effect of the dose of normal lung tissue under different fixed modes by a comparative analysis of combined body position and thermoplastic film fixed set-up error of radiation therapy for lung cancer. Methods From October 2016 to March 2018, the patients who received chest radiology at the Tangshan people's hospital were enrolled as subjects retrospectively divided into two groups, including 50 patients with lung cancer radiotherapy with combined body position fixation, and 40 patients with lung cancer with thermoplastic film fixation. The two groups of patients drew the target areas in accordance with the unified standard, and the set-up error of left and right, up and down, front and rear ( x, y, z axis) were recorded respectively after 1 time/week cone CT( CBCT) matched with the planned CT image and analyzed by t test. According to the MPTV =2. 5Σ+0. 7δ, CTV to PTV external expansion boundary in the combined body position group were calculated. And the V5、V20 and V30 of two groups of patients were calculated and analyzed by TPS system. Results The set-up error of the combined body position group and thermoplastic film group were respectively (1. 00 ± 0. 58) mm and (3. 28 ± 0. 43) mm on the x axis, (1. 42 ± 0. 28) mm on the y axis and (4. 03 ± 0. 41) mm, (1. 06 ± 0. 44) mm and (3. 18 ± 0. 34) mm on the z axis. The set-up errors of the two groups were statistically significant on x, y and z axis( t= -20. 740, -35. 596, -25. 015,P<0. 05). There was no significant difference in set-up errors between the central and peripheral lung cancer patients and between left and right lung cancer patients(P>0. 05). Through the MPTV =2. 5Σ+0. 7δ, CTV to PTV external expansion boundary in the combined body position fixation group was 2. 906 , 3. 746 and 2. 958 mm on x, y and z axis respectively. The comparison between group A and B showed that the mean values of V5 , V20 and V30 in group B were reduced by 1. 5％, 3. 1％ and 4. 8％ respectively compared with group A. Conclusions The combined body position technique can improve the accuracy of lung cancer patients after radiation therapy,and further reduce the boundary of CTV to PTV, which is of certain value to reduce the occurrence of radiation pneumonitis.

Construction of Wip1 gene recombinant lentiviral expression vector and its effects on breast cancer MCF-7 cell biological behavior / 重庆医学

Objective To construct the Wip1 gene recombinant lentiviral expression vector and to investigate its effects on breast cancer cell biological behaviors .Methods Wip1 gene short hairpin RNAs (shRNA) was transfected into breast cancer MCF-7 cells through lentiviral infection method .Wip1 mRNA and protein expressions before and after transfection were detected by u-sing qRT-PCR and Western blotting .The effects of Wip1-shRNA on the proliferation ,apoptosis ,cell cycle ,invasion and metastasis in MCF-7 cells were determined by using the MTT assay ,flow cytometry and transwell invasion test .Interfering RNA molecule p53dsRNA inhibiting p53 gene expression (p53 dsRNA) was screened ,and the effect of p53 inhibition on MCF-7 cells invasion and metastasis was analyzed .Results After transfection for 48 h ,the cellular fluorescence in the Wip1-shRNA group was stronger , while which in the NC-shRNA group was weaker .Cellular Wip1 mRNA and protein expressions in the Wip1 shRNA group were 0 .291 ± 0 .025 and 0 .203 ± 0 .021 respectively ,which in the NC-shRNA group were 0 .954 ± 0 .090 and 0 .963 ± 0 .092 respectively , the difference between the two groups was statistically significant (P<0 .05) .The cellular survival rate at various time points had statistical difference between the two groups (P<0 .05) .The early and late cell apoptosis number in the NC-shRNA group was less than that in the Wip1-shRNA group ,the difference was statistically significant (P<0 .05) .The cells numbers at phase G0 +G1 and phase S in the NC-shRNA group were 53 .5 ± 3 .6 and 27 .3 ± 1 .5 respectively ,which in the Wip-shRNA group were 72 .3 ± 5 .2 and 14 .6 ± 0 .8 respectively ,the difference was statistically significant(P<0 .05) .The Transwell invasion and metastasis results showed that the cell transmembrane number in the NC-shRNA group was more than that in the Wip1-shRNA group(P<0 .05) .The cellu-lar p53 mRNA and protein expression had statistical difference between the control group and p53dsRNA group(P<0 .05) .Conclu-sion RNA interference can effectively suppress Wip1 expression in MCF-7 cells .Wip1 may affect the proliferation ,apoptosis ,cell cycle ,invasion and metastasis of breast cancer cells by modulating protein expression .p53dsRNA increases the invasion and metas-tasis ability of breast cancer MCF-7 cells by interfering p53 gene down-regulation .

Research progress of capecitabine combined chemotherapy with biweekly scheme / 基础医学与临床

As an oral chemotherapy drug, capecitabine is safe, effective and easy to use, has been widely used in the treatment of esophageal cancer, gastric cancer, colorectal cancer, breast cancer, ovarian cancer and other solid tumor.The chemotherapy scheme is mainly for 3 weeks, each cycle of capecitabine for 14 days, and 7 days off.As a modification to conventional chemotherapy, the biweekly scheme with capecitabine showed low toxicity and well tolerated by patients, short hospitalization time and other characteristics, it is worth of application in clinical.

Circular RNA and the relation of carcinoma / 中国综合临床

Objective Cyclic RNA ( circRNAs) was discovered in 1970s,which is different from other RNA,its structure is special,which is a closed covalent ring structure,and shows high and stable expression in eukaryotic cells?In recent years, the study found that a large number of endogenous circRNAs exists in mammalian cells,and most circRNAs with stable expression,RNA enzyme degradation resistance,usually in the tissue cells with the characteristic of diversity and specificity?CircRNAs most formed by exons,part formed by introns?For the functional study of circRNAs,circRNAs has a similar competitive endogenous RNA ( ceRNA) of the sponge function,but also can regulate transcription and translation?More and more evidence indicates that circRNAs circRNAs may be abnormal expression in many diseases,especially in tumor cells?This could be some new diagnostic and predictive biomarkers of disease?CircRNAs in the field of RNA is becoming a new research hotspot,and can be widely involved in many areas of life.

Expression of BTG1 protein in laryngeal squamous cell carcinoma and its clinical significance / 临床耳鼻咽喉头颈外科杂志

OBJECTIVE@#To investigate the expression of B-cell translocation gene 1 (BTG1) and to determine the relationship between BTG1 expression and clinicopathological features, biological behaviors in laryngeal squamous cell carcinoma.@*METHOD@#Immunohistochemistry and Western blot were used to analyze BTG1 protein expression in 70 cases of laryngeal cancer and 35 cases of adjacent corresponding laryngeal mucosal tissues to illuminate the relationship between BTG1 expression and clinical factors.@*RESULT@#The positive rate of BTG1 protein expression was 31.43% in laryngeal carcinoma tissues, significantly lower than 91.43% in the adjacent laryngeal tissues (P 0.05) of patients with laryngeal cancer.@*CONCLUSION@#The expression of BTG1 protein was decreased in laryngeal squamous cell carcinoma, suggesting that BTG1 gene may be closely associated with the carcinogenesis and the degree of malignancy. Detection of BTG1 expression may be useful in diagnosis, treatment and prognosis of laryngeal carcinoma.

The effect of BTG1 overexpression on the proliferation and apoptosis of laryngeal cancer cells and its molecular mechanismin vitro / 中国癌症杂志

Background and purpose:B-cell translocation gene 1(BTG1) can inhibit cell proliferation, promote cell apoptosis and regulate cell cycle progression and differentiation in a variety of cell types. This study aimed to explore the inlfuence on cell proliferation, apoptosis and cell cycle and its related mechanism of laryngeal cancer Hep - 2 cell lines through BTG1 overexpression byin vitro experiments.Methods:The BTG1 expression plasmids were constructed and transfected into Hep-2. They were divided into experimental group (transfected BTG1 of Hep-2 cells) and control group (transfected empty plasmid of Hep-2 cells). Western blot method was used to identify BTG1 protein expression levels of cells; proliferation activity of cells was detected by MTT assay; lfow cytometry was used to analyze the cell cycle distribution and AnnexinⅤ-FITC/PI cell apoptosis; Western blot was also used to assay cell cycle regulatory protein and apoptosis-related protein expression.Results:The pEGFP-N1-BTG1 plasmid was constructed successfully, and the expression of BTG1 protein was higher in experimental group than that in control group (0.921±0.091vs 0.308±0.047,P<0.05). Compared with the two group of laryngeal cancer Hep-2 cells, the cell growth in experimental group was slowed down and the proliferation was reduced (P<0.05); Cyclin D1 protein expression level was decreased (0.436±0.023vs 0.916±0.092,P<0.05), the proportion of G0/G1 phase cell cycle was increased [(85.1±5.2)%vs (63.8±3.1)%,P<0.05], the proportion of S phase cell was decreased [(8.3±1.1)%vs (23.1±1.5)%, P<0.05], phosphatidylserine ectropion in experimental group was increased, cell early apoptosis was significant [(10.3±1.1)%vs (2.8±0.3)%,P<0.05] and anti-apoptotic protein Bcl-2 expression level was reduced(0.167±0.009vs 0.834±0.084,P<0.05).Conclusion:BTG1 high expression could inhibit the proliferation growth of laryngeal Hep-2 cells and promote its apoptosis, and the possible mechanisms are interrelated with BTG1 involved in cell cycle regulation and causing cell apoptosis.

The biological effect of Survivin siRNA on esoPhageal carcinoma cell line by increasing targeting CasPase-9 / 中国综合临床

Objective To investiGate the bioloGical effect of inhibitor of apoptosis proteins( Survivin) on esophaGeal cancer ECA-l09 cell line. Methods Survivin siRNA was transiently transfected into non small esophaGeal cancer ECA-l09 cell line by liposome-mediated method and the expression level of Survivin siRNA was detected by RT-PCR and western blot. MTT assay,cell apoptosis,cell cycle and western blot were conducted as to observe proliferation,apoptosis,cycle of cell and expression of Caspase-9. Results RT-PCR and Western blot showed that ECA-l09 cell transfected Survivin siRNA had a lower relative expressive content compared with untransfected Group( P <0. 05 ). MTT assay,cell apoptosis and cell cycles demonstrated that ECA-l09 cell transfected Survivin siRNA had a lower survival fraction,hiGher cell apoptosis,more percentaGe of the G2/M phases( P < 0. 05 ). Conclusion Survivin involve in the bioloGical processes of esophaGeal cancer cell proliferation,apoptosis,and cell cycle.

UHRF1 expression inhibition by RNA interference enhances the radiosensitivity of esophageal cancer cells / 中华放射肿瘤学杂志

Objective To study the effect of UHRF1 expression inhibition by RNA interference on the radiosensitivity of esophageal cancer cell line TE-1 and its mechanism.Methods Short hairpin RNA (shRNA) targeting UHRF1 gene was introduced into TE-1 cells by lentivector-mediated transfer.The cells were divided into three groups:non-transfected group,negative control (NC)-shRNA-transfected group,and UHRF1-shRNA-transfected group.The mRNA and protein expression levels of UHRF1 in TE-1 cells were measured by RT-PCR and Western blot before and after transfection.After transfection and X-ray radiation,the radiosensitivity of TE-1 cells was evaluated by colony formation assay; the cell cycle and cell apoptosis were determined by flow cytometry; the γ-H2AX (as a marker of DNA damage) level was measured by Western blot.Results After transfection with UHRF1-shRNA,the mRNA and protein expression levels of UHRF1 were significantly decreased in TE-1 cells,as compared with those in the NC-shRNA-transfected group and non-transfected group (0.11 vs 0.96 and 0.98,F =124.21,P =0.000;0.10 vs 0.89 and 0.94,F =125.25,P =0.000).The UHRF1-shRNA-transfected group had sensitization enhancement ratios of 1.53 (D0 ratio) and 1.95 (Dq ratio).X-ray radiation could cause G2/M arrest and increase apoptotic rate and γ-H2AX expression in TE-1 cells.Compared with the two control groups,the UHRF1-shRNA-transfected group showed significantly less G2/M arrest (F =500.15,P =0.000),a significantly higher apoptotic rate (F =100.10,P =0.000),and significantly higher residual γ-H2AX expression (F =61.00,P =0.000) at 24 hours after X-ray radiation.Conclusions RNA interference can effectively inhibit the UHRF1 expression and enhance the radiosensitivity of TE-1 cells.The mechanism may be related to cell cycle regulation,cell apoptosis,and DNA damage repair.

Relationship between Survivin protein and esophageal cancer: a systematic review / 肿瘤研究与临床

Objective To investigate Survivin protein and esophageal cancer with a systematic review.Methods The published studies were searched in the Cochrane Library (Issuel,2011),Pubmed and CNKI databases, and other relevant journals were also hand searched to identify all the relevant case-control trials.Then the quality of the included trials was assessed and meta-regression analysis was performed,by STATA 11.0 software.Results A total of 15 studies were recruited.As for the positive rate of Survivin expression, significant differences were tested between esophageal cancer vs.normal esophageal tissues (OR =39.98, 95 ％ CI =15.08-106.02, P ＜ 0.05).No significant difference was tested between cell differentiation G3 vs.cell differentiation G1-G2(OR =1.47,95 ％ CI =0.59-3.68,P ＞ 0.05).There were significant differences between T stagesⅢ-Ⅳ vs.T stages Ⅰ-Ⅱ (OR =2.82,95 ％ CI =1.35-5.90,P ＜ 0.05),clinic stages Ⅲ-Ⅳ vs.clinic stages Ⅰ-Ⅱ (OR =2.99, 95 ％ CI =2.03-4.41, P ＜ 0.05), lymph node metastasis vs.non-lymph node metastasis (OR =4.11,95 ％ CI =2.79-6.05,P ＜ 0.05).Moreover,Survivin expression was positively correlated with the expression of hypoxia inducible factor-1α (HIF-1α) (r =0.542,P ＜ 0.005).Conclusion Survivin expression increased malignant behaviors of esophageal cancer.There might be substantial correlations between Survivin and HIF-1α expression and esophageal cancer.

Para-recurrent lymph node metastasis was a significant predictor for cervical lymph nodes metastasis in thoracic esophageal carcinoma / 中华放射肿瘤学杂志

ObjectiveTo evaluate correlation factors of cervical lymph nodes metastasis in thoracic esophageal carcinoma.MethodsLocal-regional metastasis of lymph node for 126 cases with esophageal squamous cell cancer after surgery from 2004 to 2009 were reviewed.Risk factors of cervical lymph nodes metastasis were examined by multiple Logistic regression analysis.ResultsIn 126 cases,supraclavicular lymph node metastasis rate was 43.7％ (55/126).By logistic regression,none of the primary site,T stage,N stage,histological grade,lymph node metastasis rate,lymph node metastasis degree and number of lymph nodes metastatic field was not the high risk of cervical lymph nodes metastasis.In addition,multivariate analysis found that lymph node metastasis in mediastinum region 1 was high risk factor for lymph node metastasis of region 1 ( x2 =12.14,9.27,P =0.000,0.002),lymph node metastasis in region Ⅲ and region 2 were high risk factors for lymph node metastasis of region Ⅱa ( x2 =14.56,8.27,8.02,3.93,P =0.000,0.004,0.005,0.047 ).ConclusionMediastinal para-recurrent nerve lymph node metastasis is a significant predictor for cervical lymph nodes metastasis.

Recurrence patterns of thoracic esophageal cancer after two-field lymph node dissection / 中华放射肿瘤学杂志

Objective To investigate the local-regional recurrence in thoracic esophageal cancer after radical surgery including two-field lymph node dissection and provide evidence for postoperative radiotherapy. Methods We reviewed local-regional recurrence for 134 cases with esophageal squamous cell carcinoma after radical surgery from 2004 to 2009. Results In 134 cases, lymph node metastasis rate,anastomosis recurrence rate and tumor bed recurrence rate was 94. 0%, 9. 7% and 3.7%, respectively. As to the 126 cases with lymph node metastasis, significant difference was detected between mediastinal metastasis, supraclavicular metastasis and abdominal lymph node metastasis (80. 2%, 43.7% and 13.5%,respectively, χ2= 113. 15, P = 0. 000). Furthermore, the relative metastasis rate in upper mediastinum,middle mediastinum and the lower mediastinum was 73.8%, 39.7% and 1.6%, respectively, the difference was statistically significant ( χ2 = 139. 11, P = 0. 000 ). Significant difference was identified between right and left supraclavicular lymph node metastasis (31.7% vs 16. 7%, χ2= 7. 81, P = 0. 005 ).To confirm the analysis above,lymph node metastasis rate of left recurrent laryngeal nerve nodes, (including region 1L, 2L, 4L and 5) ,right recurrent laryngeal nerve nodes, azygos nodes, subcarinal nodes, and 2R region was 38.9%, 43.7%, 15.1%, 34.1% and 25.4%, respectively. Conclusions The main characteristics of local-regional recurrence may be lymph node metastasis for esophageal squamous cell carcinoma after radical surgery. On the contrary, tumor bed recurrence is rare. Dangerous regions include supraclavicular nodes, recurrent laryngeal nerve nodes, azygos nodes as well as subcarinal nodes.