Renal cell carcinoma (RCC) is one of the common malignant tumors in human beings and originates from the renal tubular epithelium. Clear cell renal cell carcinoma (ccRCC) is the main pathological type of RCC. Due to the lack of reliable biomarkers and clinical symptoms for early diagnosis, imaging findings such as ultrasound and CT are needed. When the patients presented typical symptoms, for example, hematuria, backache, and abdominal mass, some of them are in advanced stages of cancer. About a quarter of patients had metastasis at the first diagnosis, and the 5-year survival rate of these patients was less than 10%. Therefore, the early diagnosis of ccRCC and the prevention of tumor recurrence and metastasis are of great significance. The preliminary studies suggested that some hypoxia and metabolism-related molecules were highly expressed in ccRCC tumors but low in normal kidney tissues. The molecules included carbonic anhydrase IX/9 (CA IX/CA9), the mitochondrial NADH dehydrogenase \[ubiquinone\] 1 alpha subcomplex, 4-like 2(NDUFA4L2), angiopoietin-like protein 4(ANGPTL4), hypoxia inducible lipid droplet-associated (HILPDA）, and egl-9 family hypoxia-inducible factor 3( EGLN3) et al . Cell-free DNA methylomes were also highly expressed in the blood of ccRCC patients. In order to further verify the expression status of the above novel biomarkers in ccRCC, the investigators will detect the expressions of these molecules in the tumor and adjacent tissues from 140 ccRCC patients by RT-PCR, Western blot, and immunohistochemistry.140 healthy people were selected as the control group. 30 patients with benign kidney diseases were selected as another control group. Blood and urine samples from the ccRCC group and the control group were collected. The mRNA and protein levels of the above molecules in blood or urine samples were detected by qRT-PCR and ELISA. The correlation between the expression of the above new biomarkers and clinical data, such as early diagnosis, pathological grade, recurrence and metastasis, and survival time, was statistically analyzed. The above molecular changes were dynamically detected before surgery, 1 week, and 6 months after surgery. A receiver-operating characteristic curve (ROC) was used to determine the threshold value of these biomarkers for the diagnosis of renal clear cell carcinoma. The study is to explore the specific tumor biomarker spectrum for clinical diagnosis, evaluation of recurrence, metastasis, and prognosis of ccRCC, which will be auxiliary early screening and diagnosis, reducing the harm of renal cancer to human health.
Study Type
OBSERVATIONAL
Enrollment
300
The patient was treated normally and no intervention was required in this study
The First Affiliated Hospital of Shandong First Medical University & Shandong Provincial Qianfoshan Hospital
Jinan, China
RECRUITINGDetection of novel biomarkers in ccRCC patients before surgery
1. The expressions of novel biomarkers in blood samples: The protein levels of CA9, NDUFA4L2, ANGPTL4, HILPDA and EGLN3 in blood samples were detected by ELISA method. The levels of cell-free DNA methylomes in blood samples were detected by real-time fluorescence quantitative PCR method. 2. The expressions of novel biomarkers in urine samples: The levels of cell-free DNA methylomes in urine samples were detected by real-time fluorescence quantitative PCR method.
Time frame: For ccRCC patients, these six novel biomarkers were detected 1 to 3 days before surgery
Detection of novel biomarkers in ccRCC patients 1 week after surgery
1. The expressions of novel biomarkers in tissues: In the tumor and adjacent tissues from ccRCC patients, the mRNA expressions of CA9, NDUFA4L2, ANGPTL4, HILPDA and EGLN3 were measured by real-time fluorescence quantitative PCR method. The protein expressions of these five molecules were detected by western blot, and immunohistochemistry methods. 2. The expressions of novel biomarkers in blood samples: The protein levels of CA9, NDUFA4L2, ANGPTL4, HILPDA and EGLN3 in blood samples were detected by ELISA method. The levels of cell-free DNA methylomes in blood samples were detected by real-time fluorescence quantitative PCR method. 3. The expressions of novel biomarkers in urine samples: The levels of cell-free DNA methylomes in urine samples were detected by real-time fluorescence quantitative PCR method.
Time frame: For ccRCC patients, the detection of these biomarkers were completed within one week after surgery
Detection of novel biomarkers in ccRCC patients at 6 months after surgery
1. The expressions of novel biomarkers in blood samples: The protein levels of CA9, NDUFA4L2, ANGPTL4, HILPDA and EGLN3 in blood samples were detected by ELISA method. The levels of cell-free DNA methylomes in blood samples were detected by real-time fluorescence quantitative PCR method. 2. The expressions of novel biomarkers in urine samples: The levels of cell-free DNA methylomes in urine samples were detected by real-time fluorescence quantitative PCR method.
Time frame: For ccRCC patients, these six novel biomarkers were detected at 6 months postoperatively
Detection of novel biomarkers in the control groups as baseline
1. The expressions of novel biomarkers in blood samples: The protein levels of CA9, NDUFA4L2, ANGPTL4, HILPDA and EGLN3 in blood samples were detected by ELISA method. The levels of cell-free DNA methylomes in blood samples were detected by real-time fluorescence quantitative PCR method. 2. The expressions of novel biomarkers in urine samples: The levels of cell-free DNA methylomes in urine samples were detected by real-time fluorescence quantitative PCR method.
Time frame: For the healthy control group and the benign kidney disease group, the levels of these six biomarkers were detected as baseline.
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