Potential biomarkers for noninvasive diagnosis of this disease [6?]. But the frequency of DNA methylation in tumor suppressor genes between cancer tissue and autologous clinical samples ranged a lot among the published 298690-60-5 biological activity studies with small sample size. Accordingly, we performed a metaanalysis on the basis of published articles of P16INK4a promoter methylation and lung cancer in order to better identify the correlation of methylation 86168-78-7 site status between cancer tissue and autologous samples.and corresponding free text word searching term. The title and abstract of initial identified articles were evaluated for appropriateness to the inclusion criteria. Then all potentially relevant articles were assessed in full-text paper and all references of included articles were further scanned for additional analysis.Data Extraction and Quality AssessmentThe inclusion criteria of the meta-analysis was as follows: the patients were limited to non-small cell lung carcinoma without restriction 
of stages. The methods used for methylation detection were confined to methylation-specific polymerase chain reaction(MSP), real-time MSP(RT-MSP) and quantitative MSP(qMSP). The results were the P16INK4A gene promoter methylation status in tumor tissue and corresponding autologous controls, including non-tumor lung tissue(NLT), plasma, sputum and bronchoalveolar lavage fluid(BALF) of NSCLC patients. Information on the name of the first author, year of publication, region of the included subjects and methylation status of P16INK4A gene in cancer tissue and controls were recorded from each study. Detailed information about each article was extracted by two reviewers (JG and YW) and then checked by the third reviewerMaterials and Methods Studies IdentificationThe selection procedure of studies was illustrated in the PRISMA (Preferred Reporting Items for Systematic Reviews and Meta-Analyses) statement flow chart (Fig. 1). Studies about P16INK4a gene promoter methylation in NSCLC, published before January 2012, were identified through an electronic sensitive search of Medline, EMBSE and CNKI databases. The searching strategy was performed using “Non-Small-Cell Lung Carcinoma” AND “methylation” as the Medical Subject Headings (MeSH)Figure 1. PRISMA flowchart of the literature search strategy for systematic review. (Data from some studies was used more than once, as they reported data in multiple controls.). doi:10.1371/journal.pone.0060107.gP16INK4a Promoter Methylation 1662274 and NSCLCTable 1. General characteristics of included studies.Sample size (n) Author Seike [13] Su [16] He [31] Zochbauer [6] Bearzatto [30] Chen [25] He [32] Ng [17] Cai [33] Harden [26] Liu [18] Guo [27] Liu [14] Zhang [34] Russo [19] Georgiou [23] Li [35] Rosalia [29] Ulivi [15] Wang [36] Belinsky [20] Hong [22] Hsu(1) [21] Hsu(2) [24] Kim [28] Yang [37] Zhang [38] Guo [39] Wang [8] Chen [40] Guo [41] Zhang [42] Zhang [7] Sun [43] Year publication Location 2000 2000 2001 2001 2002 2002 2002 2002 2003 2003 2003 2004 2004 2004 2005 2007 2006 2006 2006 2006 2007 2007 2007 2007 2007 2007 2007 2008 2008 2010 2010 2006 2011 2012 Japan China China USA Italy Taiwan China Hong Kong China USA China USA China China USA Greece China Italy Italy China England Korea Taiwan Taiwan Korea China China China China China China China China China Age(y) 63.7(40?0) 58.9 Na Na 64 Na Na 60.2 59.5 67(40?7) Na 66.1(42?3) Na Na Na 63(38?6) Na 60.2(51?4) Na 32?3 62(37?0) Na 69 Na 6368.4 56(31?7) Na 59613 Na 59.7(32?9) 59.2 52.3(37.Potential biomarkers for noninvasive diagnosis of this disease [6?]. But the frequency of DNA methylation in tumor suppressor genes between cancer tissue and autologous clinical samples ranged a lot among the published studies with small sample size. Accordingly, we performed a metaanalysis on the basis of published articles of P16INK4a promoter methylation and lung cancer in order to better identify the correlation of methylation status between cancer tissue and autologous samples.and corresponding free text word searching term. The title and abstract of initial identified articles were evaluated for appropriateness to the inclusion criteria. Then all potentially relevant articles were assessed in full-text paper and all references of included articles were further scanned for additional analysis.Data Extraction and Quality AssessmentThe inclusion criteria of the meta-analysis was as follows: the patients were limited to non-small cell lung carcinoma without restriction of stages. The methods used for methylation detection were confined to methylation-specific polymerase chain reaction(MSP), real-time MSP(RT-MSP) and quantitative MSP(qMSP). The results were the P16INK4A gene promoter methylation status in tumor tissue and corresponding autologous controls, including non-tumor lung tissue(NLT), plasma, sputum and bronchoalveolar lavage fluid(BALF) of NSCLC patients. Information on the name of the first author, year of publication, region of the included subjects and methylation status of P16INK4A gene in cancer tissue and controls were recorded from each study. Detailed information about each article was extracted by two reviewers (JG and YW) and then checked by the third reviewerMaterials and Methods Studies IdentificationThe selection procedure 
of studies was illustrated in the PRISMA (Preferred Reporting Items for Systematic Reviews and Meta-Analyses) statement flow chart (Fig. 1). Studies about P16INK4a gene promoter methylation in NSCLC, published before January 2012, were identified through an electronic sensitive search of Medline, EMBSE and CNKI databases. The searching strategy was performed using “Non-Small-Cell Lung Carcinoma” AND “methylation” as the Medical Subject Headings (MeSH)Figure 1. PRISMA flowchart of the literature search strategy for systematic review. (Data from some studies was used more than once, as they reported data in multiple controls.). doi:10.1371/journal.pone.0060107.gP16INK4a Promoter Methylation 1662274 and NSCLCTable 1. General characteristics of included studies.Sample size (n) Author Seike [13] Su [16] He [31] Zochbauer [6] Bearzatto [30] Chen [25] He [32] Ng [17] Cai [33] Harden [26] Liu [18] Guo [27] Liu [14] Zhang [34] Russo [19] Georgiou [23] Li [35] Rosalia [29] Ulivi [15] Wang [36] Belinsky [20] Hong [22] Hsu(1) [21] Hsu(2) [24] Kim [28] Yang [37] Zhang [38] Guo [39] Wang [8] Chen [40] Guo [41] Zhang [42] Zhang [7] Sun [43] Year publication Location 2000 2000 2001 2001 2002 2002 2002 2002 2003 2003 2003 2004 2004 2004 2005 2007 2006 2006 2006 2006 2007 2007 2007 2007 2007 2007 2007 2008 2008 2010 2010 2006 2011 2012 Japan China China USA Italy Taiwan China Hong Kong China USA China USA China China USA Greece China Italy Italy China England Korea Taiwan Taiwan Korea China China China China China China China China China Age(y) 63.7(40?0) 58.9 Na Na 64 Na Na 60.2 59.5 67(40?7) Na 66.1(42?3) Na Na Na 63(38?6) Na 60.2(51?4) Na 32?3 62(37?0) Na 69 Na 6368.4 56(31?7) Na 59613 Na 59.7(32?9) 59.2 52.3(37.
