TRIP13 Enhances Radioresistance of Lung Adenocarcinoma Cells through the Homologous Recombination Pathway / 中国肺癌杂志

BACKGROUND@#Radiation therapy is one of the most common treatments for non-small cell lung cancer (NSCLC). However, the insensitivity of some tumor cells to radiation is one of the major reasons for the poor efficacy of radiotherapy and the poor prognosis of patients, and exploring the underlying mechanisms behind radioresistance is the key to solving this clinical challenge. This study aimed to identify the molecules associated with radioresistance in lung adenocarcinoma (LUAD), identified thyroid hormone receptor interactor 13 (TRIP13) as the main target initially, and explored whether TRIP13 is related to radioresistance in LUAD and the specific mechanism, with the aim of providing theoretical basis and potential targets for the combination therapy of LUAD patients receiving radiotherapy in the clinic.@*METHODS@#Three datasets, GSE18842, GSE19188 and GSE33532, were selected from the Gene Expression Omnibus (GEO) database and screened for differentially expressed genes (|log FC|>1.5, P<0.05) in each of the three datasets using the R 4.1.3 software, and then Venn diagram was used to find out the differentially expressed genes common to the three datasets. The screened differential genes were then subjected to protein-protein interaction (PPI) analysis and module analysis with the help of STRING online tool and Cytoscape software, and survival prognosis analysis was performed for each gene with the help of Kaplan-Meier Plotter database, and the TRIP13 gene was identified as the main molecule for subsequent studies. Subsequently, the human LUAD cell line H292 was irradiated with multiple X-rays using a sub-lethal dose irradiation method to construct a radioresistant cell line, H292DR. The radioresistance of H292DR cells was verified using cell counting kit-8 (CCK-8) assay and clone formation assay. The expression levels of TRIP13 in H292 and H292DR cells were measured by Western blot. Small interfering RNA (siRNA) was used to silence the expression of TRIP13 in H292DR cells and Western blot assay was performed. The clone formation ability and migration ability of H292DR cells were observed after TRIP13 silencing, followed by the detection of changes in the expression levels of proteins closely related to homologous recombination, such as ataxia telangiectasia mutated (ATM) protein.@*RESULTS@#Screening of multiple GEO datasets, validation of external datasets and survival analysis revealed that TRIP13 was highly expressed in LUAD and was associated with poor prognosis in LUAD patients who had received radiation therapy. And the results of gene set enrichment analysis (GSEA) of TRIP13 suggested that TRIP13 might be closely associated with LUAD radioresistance by promoting homologous recombination repair after radiation therapy. Experimentally, TRIP13 expression was found to be upregulated in H292DR, and silencing of TRIP13 was able to increase the sensitivity of H292DR cells to radiation.@*CONCLUSIONS@#TRIP13 is associated with poor prognosis in LUAD patients treated with radiation, possibly by promoting a homologous recombination repair pathway to mediate resistance of LUAD cells to radiation.

CTCs Detection and Whole-exome Sequencing Might Be Used to Differentiate Benign and Malignant Pulmonary Nodules / 中国肺癌杂志

BACKGROUND@#Low-density computed tomography (LDCT) improved early lung cancer diagnosis but introduces an excess of false-positive pulmonary nodules data. Hence, accurate diagnosis of early-stage lung cancer remains challenging. The purpose of the study was to assess the feasibility of using circulating tumour cells (CTCs) to differentiate malignant from benign pulmonary nodules.@*METHODS@#122 patients with suspected malignant pulmonary nodules detected on chest CT in preparation for surgery were prospectively recruited. Peripheral blood samples were collected before surgery, and CTCs were identified upon isolation by size of epithelial tumour cells and morphological analysis. Laser capture microdissection, MALBAC amplification, and whole-exome sequencing were performed on 8 samples. The diagnostic efficacy of CTCs counting, and the genomic variation profile of benign and malignant CTCs samples were analysed.@*RESULTS@#Using 2.5 cells/5 mL as the cut-off value, the area under the receiver operating characteristic curve was of 0.651 (95% confidence interval: 0.538-0.764), with a sensitivity and specificity of 0.526 and 0.800, respectively, and positive and negative predictive values of 91.1% and 30.3%, respectively. Distinct sequence variations differences in DNA damage repair-related and driver genes were observed in benign and malignant samples. TP53 mutations were identified in CTCs of four malignant cases; in particular, g.7578115T>C, g.7578645C>T, and g.7579472G>C were exclusively detected in all four malignant samples.@*CONCLUSIONS@#CTCs play an ancillary role in the diagnosis of pulmonary nodules. TP53 mutations in CTCs might be used to identify benign and malignant pulmonary nodules.

Patterns of failure after targeted therapy and importance of combined radiotherapy for advanced non-small cell lung cancer / 中华放射医学与防护杂志

Objective To analyze clinical significance of failure patterns and combined radiotherapy for advanced non-small cell lung cancer after EGFR-TKIs treatment.Methods A total of 111 patients who were treated with EGFR-TKIs for advanced non-small cell lung cancer (NSCLC) with EGFR exon mutation in Peking University Cancer Hospital from January 2009 to June 2013 were retrospectively analyzed.The impact of various failure patterns and combined radiotherapy on survial were analyzed with Kaplan-Meier method.Results Totally 111 patients were enrolled in the study.The median follow-up was 27.7 months (6.6-85.3 months).The median age,median PFS andmedian OS were 59 years (35-80 years old),10.3 months (6.2-30.5 months),and 29.8 months (7.1-90.7 months),respectively.The main failure mode was the progress of the original lesion (65 cases,58.6％) and the main failure site was the progress of intrathoracic lesions (57 cases,51.4％).The survival time of patients with oligoprogress (1-3 lesions during drug resistance) was significantly extended compared with the ones whose lesions were ≥ 4.The median OS were 32.5 months and 26.7 months,respectively (x2 =4.888,P＜0.05).For 43 patients with only intrathoracic progressed,there were 9 patients treated with radiotherapy and 34 patients treated without radiotherapy.The median PFS was 9.6 and 5.7 months,respectively.The median PFS of combined radiotherapy group was significantly prolonged (x2 =9.013,P＜0.05).And the median OS of retreatment after failure were 28.1 and 13.2 months,respectively,with no significant difference between two groups (P＞0.05).For 48 patients with oligo-progress,there were 12 patients treated with radiotherapy and 36 patients treated without radiotherapy.The median PFS were 9.6 and 4.2 months,respectively.The median PFS of the group treated with combined radiotherapy was significantly longer than that of the group without combined radiotherapy treatment (x2 =5.482,P＜0.05).And the median OS of retreatment after failure were 26.0 and 11.8 months,respectively.There was no significant difference between the two groups(P＞0.05).Conclusions Combined local radiotherapy can improve the PFS of patients who had only intrathoracic progress or oligo-progress after EGFR-TKIs treatment.Therefore,the patients whose T790 M mutation indicates negative or who are not in the position to perform coressoponding detection under the intrathoracic/oligo progress stage.The local intervention plays a very critical role for patients who have primary drug restisitance to GEN 1 EGFR-TKIs.

Expression and Clinical Significance of Cancer-derived Immunoglobulin G in Non-small Cell Lung Cancer by Bioinformatics and Immunohistochemistry / 中国肺癌杂志

BACKGROUND@#It was believed that immunoglobulin G (IgG) was synthesized only by B cells. However, in recent years, researchers have found that a variety of cancer cells can also synthesize IgG (cancer-IgG) which promote the development of tumors. This study analyzed the expression and clinical significance of cancer-IgG in non-small cell lung cancer (NSCLC), and initially explored its mechanism.@*METHODS@#The expression of IgG1 heavy chain gamma 1 (IGHG1) and cancer-IgG were detected by bioinformatics and immunohistochemistry in NSCLC; The gene set enrichment analysis (GSEA) method was used to explore the signaling pathways involved in IGHG1 regulation.@*RESULTS@#The expression level of cancer-IgG in NSCLC was significantly higher than that in normal tissues. The high expression group had a poor prognosis and was associated with clinical stage (P=0.042), T stage (P=0.044) and metastasis (P=0.007). GSEA analysis showed that IGHG1 was associated with cell adhesion, cytokine interaction and chemokine signaling pathway.@*CONCLUSIONS@#High expression of cancer-IgG in NSCLC is a poor prognosis factor, which may be related to the promotion of tumor invasion and metastasis.

Influence of enteral nutrition on nutritional status,treatment toxicities,and short-term outcomes in esophageal carcinoma patients treated with concurrent chemoradiotherapy: a prospective, multicenter,randomized controlled study (NCT 02399306) / 中华放射肿瘤学杂志

Objective To investigate the influence of enteral nutrition on body weight,nutritional status,treatment toxicities,and short-term outcomes in esophageal carcinoma patients treated with concurrent chemoradiotherapy(CCRT). Methods Eligible esophageal carcinoma patients were randomly assigned(2:1) to receive either CCRT combined with enteral nutrition (study group) or CCRT alone (control group). The primary endpoint was changes in the body weight during and after radiotherapy. The secondary endpoints were nutrition-related hematological parameters,the toxicities of chemoradiotherapy,the completion rate of treatment,and short-term outcomes. The differences was using χ2 or t-test. Results Between September 2014 and June 2017,203 patients were included in the study,consisting of 139 patients in the study group and 64 patients in the control group. Compared with the control group,the study group had significantly less body weight loss during and after radiotherapy (P<0.05) and significantly less decreases in serum albumin and hemoglobin (P<0.05),but there was no significant difference in the reduction in total lymphocyte count between the two groups (P>0.05).The study group had significantly lower incidence rates of grade ≥3 myelosuppression and infection and a significantly higher completion rate of chemoradiotherapy compared with the control group (P<0.05).The incidence of radiation pneumonitis and esophagitis showed no significant difference between the two groups (P>0.05).The study group had an insignificantly higher objective response rate than the control group (P>0.05). Conclusions For esophageal carcinoma patients treated with CCRT,enteral nutrition can reduce body weight loss during and after radiotherapy,improve nutritional status and treatment tolerance,and reduce toxicities.

Consensus and controversies on delineation of radiotherapy target volume for a patient with locally advanced non-small cell lung cancer / 中华放射肿瘤学杂志

Objective To investigate the consensus and controversies on the delineation of radiotherapy target volume for patients with locally advanced non-small cell lung cancer (LA-NSCLC).Methods Questionnaires including 15 questions on the delineation of radiotherapy target volume of NSCLC were sent to 12 radiation departments in China in November 2015.A patient with LA-NSCLC was selected by Fudan University Shanghai Cancer Center, and simulation CT images and medical history data were sent to the 12 radiation departments.Twelve radiation oncologists from the 12 radiation departments showed and explained the delineation of radiotherapy target volume of their own, and the patient was discussed by all experts in the sixth multidisciplinary summit forum of precise radiotherapy and chemotherapy for tumor and lung cancer.Results All receivers of the questionnaire answered the questions.The standard lung window width/level for the delineation of lung cancer was 800-1600/-600 to-750 HU, and the mediastinum window was 350-400/20-40 HU.Respiratory movement was measured by stimulator, 4D-CT, and stimulator+4D-CT with 2-5 mm expansion based on experience.The primary clinical target volume (CTV) was defined as gross target volume (GTV) plus 5-6 mm for squamous carcinoma/5-8 mm for adenocarcinoma.The metastatic lesion of mediastinal lymph nodes was delineated as 5 mm plus primary lesion in 6 departments and as primary lesion in another 6 departments.Of the 12 departments, 10 applied 5 mm of set-up error, 1 applied 3 mm, and 1 applied 4-6 mm.For V20 of the lungs, 10 departments defined it as<30%, 1 as<35%, and 1 as 28%.Nine departments defined the radiation dose of concurrent chemoradiotherapy (CCRT) for LA-NSCLC as 60 Gy in 30 fractions, 62.7 Gy in 33 fractions in 1 department, 50-60 Gy in 25-30 fractions in 1 department, and 60-70 Gy in 25-30 fractions in 1 department.For the delineation of target volume for the LA-NSCLC patient treated with CCRT, the primary planning target volume (PTV) was defined as GTV plus organ movement (IGTV) and set-up error (GTV→IGTV→PTV) in 3 departments, as CTV plus organ movement (ITV) and set-up error (GTV→CTV→ITV→PTV) in 8 departments, and as CTV plus set-up error/IGTV plus 5-6 mm for squamous carcinoma/5-8 mm for adenocarcinoma (CTV) and set-up error (GTV→CTV→PTV/GTV→IGTV→CTV→PTV) in 1 department.For the delineation of PTV in the mediastinal lymph node, GTV→IGTV→PTV was performed in 3 departments, GTV→CTV→ITV→PTV in 8 departments, and GTV→CTV→PTV in 1 department.For 10%-100% patients with LA-NSCLC, the radiation field needed to be replanned when 38-50 Gy was completed.There was no unified standard for the optimal standardized uptake value (SUV) of positron emission tomography (PET)-computed tomography (CT) simulation and delineation.Seven departments had applied magnetic resonance imaging (MRI) simulation and 10 departments had applied stereotactic body radiation therapy (SBRT) for the treatment of early-stage NSCLC.For the delineation of PTV for early-stage NSCLC (T1-2N0M0), GTV→IGTV→PTV was performed in 5 departments, IGTV→PTV in 3 departments, and GTV→CTV→ITV→PTV in 2 departments.In all the 12 departments, peripheral early-stage NSCLC was given 6.0-12.5 Gy/fraction, 3-12 fractions and central early-stage NSCLC was given 4.6-10.0 Gy/fraction, 5-10 fractions.The results of discussion on the delineation of target volume for the patient were as follows:respiratory movements should be measured by 4D-CT or simulator;the lung window width/level is 1600/-600 HU and the mediastinal window width/level is 400/20 HU;the primary controversy is whether the involved-field irradiation or elective nodal irradiation should be used for the delineation of CTVnd in the mediastinal lymph node.Conclusions Basic consensus is reached for the delineation of target volume in LANSCLC in these aspects:lung window width/level, respiratory movements and set-up error, primary lesion delineation, the radiation dose in CCRT, and the optimal time for replanning the radiation field.There are controversies on the optimal SUV in the delineation of target volume based on PET-CT simulation, the optimal dose fractionation in SBRT for early-stage NSCLC, and the delineation of CTVnd.

Research advances in radiotherapy combined with immunotherapy for cancer / 中华放射肿瘤学杂志

Radiotherapy not only acts as an important local treatment of cancer, but also plays a key role in regulation of immune function. Radiotherapy regulates anti?tumor immune responses by promoting the generation of neoantigens, regulating the release of cytokine, and enhancing the sensitivity of tumor to cell?mediated immunity. Recently, several studies and clinical practice reported the abscopal effect in some patients undergoing radiotherapy combined with immunotherapy, which showed partial or complete response of metastases outside the irradiation field, suggesting the combination therapy as a promising strategy. However, further studies are needed for the understanding of the mechanism and influencing factors for immunity such as radiation dose and fractionation scheme. This paper reviews the research advances in the mechanism of radiotherapy?immunotherapy interaction and the combination therapy for cancer.

Role of large-diameter MRI simulation in target volume delineation in radiotherapy for nasopharyngeal carcinoma / 中华放射肿瘤学杂志

Objective To investigate the role of large-diameter magnetic resonance imaging (MRI) simulation in target volume delineation in radiotherapy for nasopharyngeal carcinoma (NPC).Methods Eighteen patients with NPC underwent computed tomography (CT) simulation and MRI simulation scans and localization in the same body position,with SOMATOM Sensation Open 40-row 82-cm large-diameter CT simulator and Siemens 3T MRI MAGNETOM Skyra 70-cm large-diameter simulator,respectively.The gross tumor volume (GTV) and parotid glands were delineated on all images according to the ICRU Report 50/62,and MRI was applied to observe the changes in GTV and parotid volume during radiotherapy.Paired t-test was applied to analyze the differences between GTVCT and GTVMRI and between GTVnx-CT and GTVnx-MRI.Results GTVMRI decreased significantly compared with GTVCT,and the average volume decreased from (213.64±84.59) cm3 to (199.68±84.69) cm3(p=0.006).As for the volume of primary lesions in the nasopharynx,GTVnx-MRI was significantly smaller than GTVnx-CT,and the volume decreased from (95.75± 24.76) cm3 to (88.12±26.25) cm3 (P =0.001);as for the volume of cervical lymph nodes,GTVnd-MRI was significantly smaller than GTVnd-CT,and the volume decreased from (117.89± 72.69) cm3 to (111.56± 70.69) cm3 (P=0.018).The targets delineated by CT and MRI did not overlap completely,with major differences in skull base bone and cervical soft tissue.The volume of both parotid glands delineated on MRI image was higher than that delineated on CT image,with a major difference in the deep lobe.MRI showed that GTV was reduced by 82.64± 16.87％ during radiotherapy,and the volumes of the left and right parotid glands were reduced by (32.7± 23.95) ％ and (34.7± 21.72) ％,respectively.Conclusions The delineation of target volume based on MRI simulation is more accurate than that based on CT simulation and can achieve a smaller volume range,which helps to guide target volume delineation in radiotherapy for NPC accurately.

Prediction of GTV/LV for radiation pneumonitis in patients with stage Ⅲ non-small cell lung cancer after radical radiotherapy / 中华放射医学与防护杂志

Objective To analyze relevant clinical and dosimetric factors associated with radiation pneumonitis in patients with stage Ⅲ non-small cell lung cancer after they received radical radiotherapy.Methods A total of 126 patients with stage Ⅲ non-small cell lung cancer who received precision radiotherapy in Peking University Cancer Hospital were analyzed from January 2013 to December 2014.Data were collected including various clinical factors (including sex,age,histological type,tumor location,history of diabetes,history of hypertension,history of smoking,the season patients received treatment,ECOG performance status before treatment,chemotherapy before radiotherapy,concurrent chemotherapy and the classification of radiation pneumonitis),as well as related dosimetric parameters [including GTV,lung volume (LV),bilateral V5,V10,V20,V30 and MLD].SPSS 19.0 software was used to analyze the relation between correlation factors and radiation pneumonitis (RP≥2).Results Among the patients,31 cases (24.6％) had occurrance of radiation pneumonitis ≥ 2.Univariate analysis showed that age,ECOG performance status before treatment,concurrent chemotherapy and GTV/LV ratio were significantly correlated with RP ≥ 2 (R =0.157-0.222,P ＜ 0.05).Further multivariate Logistic regression showed that age,concurrent chemotherapy and GTV/LV ratio were significantly correlated (Wald =4.754,6.422,14.79,P ＜ 0.05).Conclusions In patients with stage Ⅲ non-small cell lung cancer after receiving thoracic radical radiotherapy,increasing age and GTV/LV ratio≥3.2％ are risk factors of RP≥2.The concurrent chemotherapy with low-dose paclitaxel might also increase the risk of RP≥2.

Association between p53 codon 72 polymorphism and curative ef-fect of postoperative chemo-radiotherapy for breast cancer / 中国肿瘤临床

Objective:To investigate the association between p53 codon 72 polymorphism and the prognosis of breast cancer pa-tients receiving chemotherapy and radiotherapy after surgery. Methods:A total of 427 breast cancer patients treated with chemo-radio-therapy after surgery at Beijing Cancer Hospital were selected for this study. Polymerase chain reaction–restriction fragment length polymorphism was adopted to analyze the p53 codon 72 polymorphism. Survival analysis was conducted to compare the disparities of recurrence and survival among the patients with different p53 codon 72 polymorphic variants. Results:The distribution of three geno-types of p53 codon 72 in our cohort is as follows:Pro/Pro 18.3%(78/427), Pro/Arg 44.0%(188/427), and Arg/Arg 37.7%(161/427). No significant difference was observed among the local recurrence-free survival (LRFS), loco-regional recurrence-free survival (LR-RFS), distant disease-free survival (DDFS), and overall survival (OS) among the three genotypes (all P>0.05). Among the 303 estro-gen receptor (ER)-positive patients, OS was significantly better in patients with Arg/Arg genotype than those with Pro/Pro genotype (χ2=6.33, P=0.042). The multivariate analysis showed that the p53 codon 72 polymorphism is an independent factor of prognosis for LRFS, LRRFS, DDFS, and OS of ER-positive patients. For the ER positive patients with Pro/Pro genotype, the local recurrence, local-regional recurrence, distant metastasis, and mortality risks were 5.9 (HR=5.9, 95%CI 1.1-31.1, P=0.036), 3.1 (HR = 3.1, 95% CI 1.1-9.1, P=0.039), 2.8 (HR=2.8, 95% CI 1.3-6.0, P=0.010), and 4.0 (HR=4.0, 95% CI 1.3-12.0, P=0.013) times higher than those with Arg/Arg genotype, respectively. Conclusion:For ER-positive breast cancer patients who underwent surgery and chemo-radiotherapy, the local recurrence, loco-regional recurrence, distant metastasis, and mortality risk with Pro/Pro genotype are significantly higher compared to those with Arg/Arg genotype.

Comparisons of two chemoradiotherapy regimens for locally advanced non-squamous and non-small cell lung cancer / 实用肿瘤学杂志

[Abstrct] Objective To compare the short -term tumor response and adverse events of two chemoradio-therapy regimens for locally advanced non -squamous non-small cell lung cancer .Methods From March 2009 to January 2013 ,we recruited 42 patients with stage ⅢNSCLC who had received three -dimensional intensity -modulated radiotherapy combined with chemotherapy .group A treated with radiotherapy combined pemetrexed plus cisplatin and group B treated with radiotherapy combined docetaxel plus cisplatin .We compared the short -term tumor response and adverse events between the two regimens .Results There were 28 cases in group A and 14 cases in group B .There were no significant differences between the two groups in hematologic toxicities ,such as leukopenia,neutropenia,anemia and thrombocytopenia.In non-hematologic toxicities,radiation pneumonitis and cough were no significances respectively (P<0.05).There were no significant differences in other non -he-matologic toxicities,such as liver dysfunction,renal dysfunction,fever,dyspnea,radiation esophagitis,hypody-namia,weight loss,gastrointestinal reactions and skin reactions .The response rate displayed no differences be-tween two groups .Conclusion This study reveals Pemetrexed plus cisplatin group had less non -hematologic toxicities than Docetaxel plus cisplatin group in locally advanced non -squamous non -small cell lung cancer . But there are no differences in the short -term tumor response between the two regimens .

Dosimetric comparison between RapidArc and fixed gantry dynamic IMRT for central-type lung cancer radiotherapy / 中华放射医学与防护杂志

Objective To compare the dosimetric difference between RapidArc and fixed gantry angle dynamic IMRT (dIMRT) for central-type lung cancer radiotherapy. Methods Therapy for 10 patients previously treated with dIMRT was replanned with RapidArc. Dose prescription was 66 Gy/33 fraction. Comparative endpoints were planning target volume (PTV) dose, doses to surrounding structures,number of monitor units, and treatment delivery time. Results There was no significant dosimetric difference between RapidArc and dIMRT. Compared with dIMRT, RapidArc slightly elevated target volume dose, lung V5, V10. The average values of lung V20, V30 and heart V30 were larger in dIMRT than those in RapidArc. The number of monitor units was reduced by 32% and the treatment time by 66% in RapidArc.Conclusions Both RapidArc and dIMRT plans could meet the clinical therapy needs. RapidArc could achieve similar target coverage and sparing of organs at risk, with fewer monitor units and shorter delivery time than dIMRT.

Consensus and controversies on delineation of radiotherapy target volume for patients with non-small cell lung cancer / 中华放射肿瘤学杂志

Objective To investigate the consensus and controversies on delineation of radiotherapy target volume for patients with non-small cell lung cancer(NSCLC). Methods Study questionnaires were designed by radiation oncologists in Peking University School of Oncology. The forms were sent through email to radiation oncologists in 10 radiation departments in China and 2 departments in US in November,2007. The brief introduction and PET/CT digital data of one patient with NSCLC were sent to radiation oncologists in 10 departments in Beijing. On Jan. 12,2008,the case discussion was held by more than 300 radiation oncologists from Beijing,Tianjin, Hebei, Liaoning Province and Inner Mongolia Autonomous Region. Results All receivers of the questionnaire responded . The set up error was 5-7 mm . For patients with locally advanced NSCLC treated with radiotherapy concurrently with near full dose chemotherapy,ll out ot 12 responding departments defined planning target volume(PTV) of primary tumor as gross tumor volume(GTV) plus 6-8 nun plus set-up error and respiratory movements ,and only one defined PTV as GTV plus set-up error and respiratory movements. For PTV of the mediastinal lymph nodes in the same patient,9 out of 12 responding departments defined PTV as GTV plus 6-8 mm plus set-up error and respiratory movements,and 3( of China) out of 12 defined PTV as GTV plus set-up error and respiratory movements. Stereotactic body .radiotherapy with high fraction dose was used in 11 out of 12 responding departments with fraction dose varying from 6 to 20 Gy,including 6 of which defined PTV of primary tumor as GTV plus 6-8 mm plus respiratory movements and set-up error, and 5 defined PTV of early stage lung cancer as GTV plus respiratory movements and set-up error. The consensus on delineation of primary tumor of the case discussion was that the appropriate window width and window level were 1600 Houasfield Units(HU) and -600 HU for lung window,and 400 HU and 20 HU for mediastinal window. The controversies was focused on whether the CTV for metastatic lymph nodes should be restricted as GTV plus 6-8 mm or enlarged to enclose all the involved lymph node region. Conclusions PIT of primary tumor and mediastinal metastatic lymph nodes should be GTV plus 6-8 mm plus respiratory movements plus set-up error. The basic controversies of target delineation are focused on the fraction dose and PIT range for early stage NSCLC, and on the possibility of defining the PIT as GTV plus respiratory movements and set-up error when treated with concurrent radiotherapy and full dose chemotherapy for locally advanced NSCLC.

Detection of CK19 and CEA mRNA expression for the diagnosis of peripheral blood micrometastases in patients with non-small cell lung cancer / 中国肺癌杂志

<p><b>BACKGROUND</b>To explore the sensitivity and clinical significance of detection of CK19 mRNA and CEA mRNA expression for the diagnosis of micrometastases in the peripheral blood of patients with non small cell lung cancer (NSCLC).</p><p><b>METHODS</b>Nested reverse transcription-polymerase chain reaction (RT-PCR) was used to detect CK19 mRNA and CEA mRNA in the peripheral blood of 35 patients with NSCLC. Twenty patients with pulmonary benign lesions and 20 normal healthy volunteers were served as controls.</p><p><b>RESULTS</b>The positive rates of CK19 mRNA and CEA mRNA for NSCLC patients were 40% (14/35) and 57% (20/35) respectively, 14 cases (40%) were positive for both mRNA, and 15 cases (43%) were both negative for both mRNA. No blood samples from patients with pulmonary benign lesions was positive for CEA mRNA, but 2 samples (10%) were positive for CK19 mRNA. Of these 20 samples from normal volunteers, none was positive for both CK19 mRNA and CEA mRNA. The distant metastasis rate of patients with positive CK-19 mRNA , positive CEA mRNA and both negative in the peripheral blood were 50%, 56% and 27% respectively.</p><p><b>CONCLUSIONS</b>Both CK-19 mRNA and CEA mRNA are good markers to detect micrometastasis in the peripheral blood of patients with NSCLC. Detection of CK19 and CEA mRNA expression might be helpful to predict the prognosis of patients with lung cancer.</p>

Detection and quantitation of CK19 mRNA in the peripheral blood and regional lymph nodes of lung cancer patients / 中国肺癌杂志

<p><b>BACKGROUND</b>To detect the micrometastases status in peripheral blood and regional lymph nodes of lung cancer patients by reverse transcription-polymerase chain reaction (RT-PCR).</p><p><b>METHODS</b>CK19 mRNA expression in peripheral blood and regional lymph nodes was detected in 78 patients with lung cancer, and 30 patients with pulmonary benign lesions and 10 healthy volunteers as controls by RT-PCR. Meanwhile, all lymph nodes were also examined by traditional pathological method.</p><p><b>RESULTS</b>The positive rate of CK19 mRNA expression was 38.5% in peripheral blood of lung cancer patients, and 6.7% in patients with pulmonary benign lesions (6.7%) (Chi-square=10.505,P=0.001). No positive CK19 mRNA expression was found in peripheral blood of 10 healthy volunteers. The positive rates of CK19 mRNA of lymph nodes were 36.9% and 0 in lung cancer patients and pulmonary benign disease patients respectively (Fisher's exact=0.014). In lung cancer group, the metastatic rate of lymph nodes was 17.9% by traditional pathological examination, which was much lower than that by RT-PCR (Chi-square=7.664, P=0.006).</p><p><b>CONCLUSIONS</b>RT-PCR amplification of CK19 mRNA is an sensitive method to detect early haematogenous and regional lymph nodes dissemination of cancer cells for patients with lung cancer. This method may lead to an earlier diagnosis and treatment of patients with subclinical metastasis in circulation and regional lymph nodes.</p>

Clinical significance of LUNX mRNA, CK19 mRNA, CEA mRNA expression in detecting micrometastasis from lung cancer / 中华放射肿瘤学杂志

Objective To evaluate the sensitivity, specificity and clinical significance of CK19 mRNA, CEA mRNA and LUNX mRNA for detecting micrometastasis by sampling the peripheral blood and regional lymph nodes of lung cancer patients. Methods Reverse transcriptase polymerase chain reaction (RT-PCR) was used to detect LUNX mRNA,CK19 mRNA, CEA mRNA for micrometastasis by sampling the peripheral blood of 48 lung cancer patients and 44 regional lymph nodes of such patients treated by curative resection. Peripheral blood of 30 patients with pulmonary benign lesions and 10 normal healthy volunteers and lymph nodes of 6 patients with benign pulmonary diseases served as control. Results (1) LUNX mRNA, CK19 mRNA, CEA mRNA were expressed in all(35/35) lung cancer tissues. (2) In the peripheral blood from 48 lung cancer patients, 30(62.5%) were positive for LUNX mRNA, 24 (50.0% ) positive for CK19 mRNA and 32(66.7%) positive for CEA mRNA. The positive detection rates of micrometastasis in 44 lymph nodes from lung cancer patients were 36.4% (16 out of 44) for LUNX mRNA, 27.3%(12 out of 44) for CK19 mRNA and 40.9%(18 out of 44) for CEA mRNA. (3) In the 30 blood samples from patients with pulmonary benign diseases, 2 (6.7%) expressed CK19 mRNA, but none expressed LUNX mRNA or CEA mRNA. All the 3 molecular markers were negative in the 10 blood samples from healthy volunteers. In 11 lymph nodes from patients with pulmonary benign lesions, none was positive for any of the three markers.(4)In 44 regional lymph nodes from lung cancer patients,6(13.6%) were positive for metastasis by histopathological examination, with a positive rate significantly lower than that of the RT-PCR ( P