Background
Methods
Quantitative analysis of RAS p21
Number of controls | 48 |
---|---|
Mean | 200,88 pg/ml |
Median | 161,00 pg/ml |
Range | 58–646 pg/ml |
Standard deviation | 125,56 pg/ml |
Mean + 2 SD | 452 pg/ml |
Detection of other biomarkers
Statistical analysis
Item to be reported | Page no. | |
---|---|---|
INTRODUCTION | ||
1 | State the marker examined, the study objectives, and any pre-specified hypotheses. | 5 |
MATERIALS AND METHODS | ||
Patients
| ||
2 | Describe the characteristics (e.g., disease stage or co-morbidities) of the study patients, including their source and inclusion and exclusion criteria. | 6 |
3 | Describe treatments received and how chosen (e.g., randomized or rule-based). | 6 |
Specimen characteristics
| ||
4 | Describe type of biological material used (including control samples) and methods of preservation and storage. | 6 |
Assay methods
| ||
5 | Specify the assay method used and provide (or reference) a detailed protocol, including specific reagents or kits used, quality control procedures, reproducibility assessments, quantitation methods, and scoring and reporting protocols. Specify whether and how assays were performed blinded to the study endpoint. | 6–7 |
Study design
| ||
6 | State the method of case selection, including whether prospective or retrospective and whether stratification or matching (e.g., by stage of disease or age) was used. Specify the time period from which cases were taken, the end of the follow-up period, and the median follow-up time. | 6 |
7 | Precisely define all clinical endpoints examined. | 7 |
8 | List all candidate variables initially examined or considered for inclusion in models. | 9 |
9 | Give rationale for sample size; if the study was designed to detect a specified effect size, give the target power and effect size. | 6 |
Statistical analysis methods
| ||
10 | Specify all statistical methods, including details of any variable selection procedures and other model-building issues, how model assumptions were verified, and how missing data were handled. | 7 |
11 | Clarify how marker values were handled in the analyses; if relevant, describe methods used for cutpoint determination. | 6 Tab. 1 |
RESULTS | ||
Data
| ||
12 | Describe the flow of patients through the study, including the number of patients included in each stage of the analysis (a diagram may be helpful) and reasons for dropout. Specifically, both overall and for each subgroup extensively examined report the numbers of patients and the number of events. | 8 Fig. 1 |
13 | Report distributions of basic demographic characteristics (at least age and sex), standard (disease-specific) prognostic variables, and tumor marker, including numbers of missing values. | 8 |
Analysis and presentation
| ||
14 | Show the relation of the marker to standard prognostic variables. | 8–9 Tab. 2 |
15 | Present univariable analyses showing the relation between the marker and outcome, with the estimated effect (e.g., hazard ratio and survival probability). Preferably provide similar analyses for all other variables being analyzed. For the effect of a tumor marker on a time-to-event outcome, a Kaplan-Meier plot is recommended. | 8 Fig. 2a,b |
16 | For key multivariable analyses, report estimated effects (e.g., hazard ratio) with confidence intervals for the marker and, at least for the final model, all other variables in the model. | 9 Tab. 3 |
17 | Among reported results, provide estimated effects with confidence intervals from an analysis in which the marker and standard prognostic variables are included, regardless of their statistical significance. | 8–9 |
18 | If done, report results of further investigations, such as checking assumptions, sensitivity analyses, and internal validation. | |
DISCUSSION | ||
19 | Interpret the results in the context of the pre-specified hypotheses and other relevant studies; include a discussion of limitations of the study. | 10–13 |
20 | Discuss implications for future research and clinical value. | 10–13 |
Results
Patients’ characteristics
RAS p21 | |||
---|---|---|---|
Total | RAS p21 elevated n (%) | p-value | |
Overall | 251 | 29 (12%) | |
ER status | 0.611 | ||
Negative | 76 | 10 (13%) | |
Positive | 174 | 19 (11%) | |
PR status | 0.358 | ||
Negative | 101 | 14 (14%) | |
Positive | 149 | 15 (10%) | |
HER2 status | 0.873 | ||
Negativea | 143 | 18 (13%) | |
Positiveb | 76 | 9 (12%) | |
Tumor subtype | 0.728 | ||
Triple-negative | 37 | 6 (16%) | |
HR-positive HER2-negative | 106 | 12 (11%) | |
HER2-positive | 76 | 9 (12%) | |
Metastatic site | 0.482 | ||
Visceral | 98 | 13 (13%) | |
Bone | 35 | 2 (6%) | |
Both | 118 | 14 (12%) | |
Extent of metastatic disease | 0.768 | ||
One site | 84 | 9 (11%) | |
Multiple sites | 167 | 20 (12%) | |
Therapeutic setting | 0.249 | ||
1st-line | 98 | 8 (8%) | |
2nd-line | 66 | 11 (17%) | |
3rd-line or more | 86 | 10 (12%) | |
Grading | 0.604 | ||
G1 | 5 | 1 (20%) | |
G2 | 129 | 13 (10%) | |
G3 | 103 | 14 (14%) | |
Circulating tumor cells | 0.101 | ||
< 5 CTCs / 7.5 ml | 122 | 17 (14%) | |
≥ 5 CTCs / 7.5 ml | 121 | 9 (7%) |
Chemotherapy | Endocrine therapy | Trastuzumab | |
---|---|---|---|
ER status | |||
Negative | 63 / 76 (83%) | 3 / 76 (4%) | 20 / 76 (26%) |
Positive | 135 / 174 (78%) | 50 / 174 (29%) | 35 / 174 (20%) |
PR status | |||
Negative | 81 / 101 (80%) | 13 / 101 (13%) | 27 / 101 (27%) |
Positive | 117 / 149 (79%) | 40 / 149 (27%) | 28 / 149 (19%) |
HER2 status | |||
Negative | 112 / 143 (78%) | 34 / 143 (24%) | 3 / 144 (2%) |
Positive | 66 / 76 (87%) | 7 / 76 (9%) | 44 / 77 (57%) |
Circulating tumor cells | |||
≥ 5 CTCs / 7.5 ml | 100 / 122 (82%) | 25 / 122 (21%) | 19 / 122 (16%) |
< 5 CTCs / 7.5 ml | 94 / 123 (76%) | 26 / 123 (21%) | 34 / 123 (28%) |
RAS p21 | |||
Elevated | 25 / 29 (86%) | 2 / 29 (7%) | 9 / 29 (31%) |
Non elevated | 174 / 222 (78%) | 51 / 222 (23%) | 46 / 222 (21%) |
RAS p21 detection
Univariate survival analysis
Multivariate survival analysis
p-value | Hazard Ratio | 95%-Confidence Interval | |
---|---|---|---|
RAS p21 Elevated vs. non elevated | 0.003 | 2.927 | 1.43–5.99 |
CTC counts ≥5 vs. < 5 CTCs / 7.5 ml blood | < 0.001 | 3.775 | 2.01–7.10 |
Therapy line > 1st line vs. 1st line | 0.002 | 2.817 | 1.48–5.37 |
Grading G3 vs. G1/2 | 0.026 | 1.380 | 1.04–1.83 |
Serum HER2 Elevated vs. non elevated | 0.156 | 1.564 | 0.84–2.90 |
Menopausal status Post- vs. Premenopausal | 0.298 | 0.732 | 0.41–1.32 |
ER status Positive vs. Negative | 0.067 | 0.478 | 0.22–1.05 |
PR status Positive vs. Negative | 0.486 | 1.297 | 0.62–2.70 |
HER2 status Positive vs. Negative | 0.121 | 0.623 | 0.34–1.13 |
Number of metastatic sites Multiple vs. Single site | 0.521 | 1.262 | 0.62–2.57 |
Metastatic spread Visceral (+/−) vs. bone only | 0.567 | 1.566 | 0.33–7.54 |
CAIX Elevated vs. non elevated | 0.300 | 1.381 | 0.75–2.54 |
TIMP1 Elevated vs. non elevated | 0.066 | 1.741 | 0.96–3.15 |