CN103278634B - Application of CD73 as stem cell surface marker of renal clear cell carcinoma - Google Patents
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Abstract
The invention discloses an application of CD73 as a stem cell surface marker of renal clear cell carcinoma. The application is that a substance for detecting CD73 can be used for preparing the following products: 1) a product for detecting or screening the stem cells of renal clear cell carcinoma; 2) a product for diagnosis or auxiliary diagnosis of renal clear cell carcinoma; 3) a product for diagnosis or auxiliary diagnosis of the malignancy degree of renal clear cell carcinoma of a patient to be detected; and 4) a product for treating renal clear cell carcinoma by taking CD73 as a target. In-vitro experiments and animal experiments prove that the CD73 can be used as a surface marker of the stem cells of renal clear cell carcinoma; and the result of clinical sample analysis indicates that the proportion of CD73 positive cells is closely related to the clinical classification of renal clear cell carcinoma. The invention provides a new molecular target CD73 to the identification of the stem cells of renal clear cell carcinoma as well as the diagnosis, classification and detection of renal clear cell carcinoma, and provides a simple and efficient method for the prognosis of renal clear cell carcinoma.
Description
Technical field
The present invention relates to the application of CD73 as stem cell surface marker of renal clear cell carcinoma.
Background technology
Kidney is also called clear-cell carcinoma, originates from uriniferous tubule epithelium.Kidney accounts for adult malignancies's 80%-90%.Usually clear-cell carcinoma is divided into 4 types: clear cell type, granular cell type, mixed cell type, neoblast type.Wherein, clear cell carcinoma of kidney (clear cell renal cell carcinoma, CCRCC) 70%-80% of kidney is accounted for, its cancer cell is often arranged in sheet, streak, acinus shape or tubulose, the spitting image of renal tubule, early stage often asymptomatic, or only have heating, the constitutional symptom such as weak, gross tumor volume is just found (J.C.Cheville when increasing, et al.Comparisons of outcome and prognostic features among histologic subtypes of renal cell carcinoma.The American journal of surgical pathology 27 (2003) 612-624).The incidence of disease of clear-cell carcinoma is growing steadily in the past 30 years always, for most of current methods for the treatment of as chemotherapy and radiation therapy, clear-cell carcinoma has resistance (L.J.Costa, et al.Renal cell carcinoma:new developments in molecular biology and potential for targeted therapies.Oncologist, 12 (2007): 1404-1415).
Tumor stem cell (tumor stem cell, TSC) is the cell having self-renewal capacity in tumour and can produce heterogeneous cell, and it is to survival, propagation, the transfer of tumour and recur important role.In essence, tumor stem cell maintains the vitality of tumor cell group by self and infinite multiplication.Motion and the ability of migrating of tumor stem cell make again the transfer of tumour cell become possibility.Tumor stem cell can be in dormant state for a long time and have several drug resistance molecule and insensitive to the extraneous chemical factors of killing tumor cell, therefore, and tumour often a period of time recurrence after conventional tumor therapeuticing method eliminates most of Common tumors cell.The qualification of tumor stem cell and therapeutic intervention are the New Policies (T.Reya, et al.Stem cells, cancer, and cancer stem cells.Nature, 414 (2001): 105-111) of the treatment of tolerance tumour.
CD73 be born of the same parents outer-5'-NT (Ecto-5 '-nucleotidase, eNT), be anchored to a kind of glycoprotein of plasma membrane by glycosyl-phosphatidyl inositol (GPI).CD73 is extensively distributed in human tissue cell surface, has hydrolytic enzyme activities and the effect of non-hydrolytic enzyme.At present, there is not yet about utilizing CD73 to carry out the relevant report of dryness analysis and clinical classification to clear cell carcinoma of kidney.
Summary of the invention
The object of this invention is to provide the application of CD73 as stem cell surface marker of renal clear cell carcinoma.
Described be applied as detect CD73(and born of the same parents outer-5'-NT) material be prepared as follows 1)-4) and in application at least one product:
1) product of clear cell carcinoma of kidney stem cell is detected;
2) product of diagnosis or auxiliary diagnosis clear cell carcinoma of kidney;
3) product of diagnosis or auxiliary diagnosis patient's clear cell carcinoma of kidney to be measured grade malignancy;
4) take CD73 as the product for the treatment of clear cell carcinoma of kidney of target spot.
Described detection CD73(and born of the same parents outer-5'-NT) material specifically can be the monoclonal antibody and related reagent and checkout equipment that detect CD73.
In above-mentioned application, described product 2) described in grade malignancy can be divided into G1, G2 and G3 tri-grades;
Described G1 level refers to that patient's clear cell carcinoma of kidney to be measured is limited in kidney peplos;
Described G2 level refers to that patient's clear cell carcinoma of kidney to be measured wears out kidney peplos, invades fat deposit, and is still confined in fascia renalis;
Described G3 level refers to that patient's clear cell carcinoma of kidney to be measured has invaded renal vein or/and inferior caval vein, invades adjacent organ or DISTANT METASTASES IN occurs.
In above-mentioned application, described product 2) comprise the carrier being described below diagnostic criteria: when CD73 positive cell ratio in patient's nephridial tissue cell to be measured is 0.49% ± 0.08%(that is 0.41-0.57%) time, be described G1 level by described patient's clear cell carcinoma of kidney candidate to be measured; When in patient's nephridial tissue cell to be measured, CD73 positive cell ratio is that 0.69% ± 0.12%(is namely 0.57%-0.81%) time, be described G2 level by described patient's clear cell carcinoma of kidney candidate to be measured; When CD73 positive cell ratio in patient's nephridial tissue cell to be measured is 1.10% ± 0.29%(that is 0.81%-1.39%) or higher than 1.39% time, be described G3 level by described patient's clear cell carcinoma of kidney candidate to be measured.
Described CD73 positive cell is the cell containing CD73.
The invention provides a kind of product detecting clear cell carcinoma of kidney stem cell, this product comprises the material of described detection CD73.
The present invention also provides the product of a kind of diagnosis or auxiliary diagnosis patient's clear cell carcinoma of kidney to be measured grade malignancy, and this product comprises the material of described detection CD73;
Described grade malignancy can be divided into G1, G2 and G3 tri-grades;
Described G1 level refers to that patient's clear cell carcinoma of kidney to be measured is limited in kidney peplos;
Described G2 level refers to that patient's clear cell carcinoma of kidney to be measured wears out kidney peplos, invades fat deposit, and is still confined in fascia renalis;
Described G3 level refers to that patient's clear cell carcinoma of kidney to be measured has invaded renal vein or/and inferior caval vein, invades adjacent organ or DISTANT METASTASES IN occurs.
This product also can comprise the described carrier recording described diagnostic criteria.
Above-mentioned CD73 specifically can be No. GenBank protein being AAH65937.1.
The present invention is proved by experiment in vitro and zoopery, and CD73 is the surface marker of clear cell carcinoma of kidney stem cell; Clinical sample analysis result shows, the clinical classification of CD73 positive cell ratio and clear cell carcinoma of kidney is closely related.The present invention is the diagnosis of the qualification of clear cell carcinoma of kidney stem cell and clear cell carcinoma of kidney, somatotype and detection provide a new molecular target CD73, and provides a simple effective method for the prognosis of clear cell carcinoma of kidney.
Accompanying drawing explanation
Fig. 1 be 786-O micro-capsule cell when free serum culture 3,7 and 14 days Electronic Speculum figure of (from left to right).Length of the scale is wherein 100 μm.
Fig. 2 is the tumour that 786-O stem cell injection NOD/SCID mouse is formed for 120 days afterwards.Place shown in left arrow is the tumour formed after injection 786-O stem cell, and place shown in right side arrow is not for forming tumour after injection 786-O Nostoc commune Vanch cell (contrast).
Fig. 3 is the CD73 protein content result of Western hybridization check 786-O stem cell.Wherein, the result of the first behavioral value CD73 albumen, the result of the second behavioral value contrast β-actin albumen; The swimming lane of left column is the testing result of common 786-O cell, and the swimming lane of right row is the testing result of 786-O stem cell.
Fig. 4 is that immunofluorescence proves that 786-O stem cell surface exists CD73 albumen.
Fig. 5 is the result of flow cytomery 786-O stem cell.Wherein, horizontal ordinate is fluorescence intensity, and ordinate is cell proportion (%).
Fig. 6 is the tumour that CD73 positive cell injection NOD/SCID mouse is formed for 120 days afterwards.Place shown in left arrow is not for forming tumour after injection CD73 negative cells, and place shown in right side arrow is the tumour formed after injection CD73 positive cell.
Fig. 7 is free serum culture CD73 negative cells and positive cell.
Fig. 8 is that the micro-capsule of CD73 negative cells and positive cell forms ration statistics result.
Fig. 9 is the SABC testing result of different clinical sample nephridial tissue section.Wherein, the result of normal kidney tissue, G1 level nephridial tissue, G2 level nephridial tissue and G3 level nephridial tissue is followed successively by from left to right.
Embodiment
The experimental technique used in following embodiment if no special instructions, is conventional method.
Material used in following embodiment, reagent etc., if no special instructions, all can obtain from commercial channels.
People's clear cell carcinoma of kidney clone 786-O: purchased from the American Type Culture Collection council of Chinese Academy of Sciences cell bank, it is numbered TCHu186.
PRMI1640 nutrient culture media: purchased from Gibco, products catalogue is numbered 12633012.
Serum: purchased from HyClone, products catalogue is numbered SV30087.02.
Pancreatin: purchased from Sigma, products catalogue is numbered T3924-500ML.
Embodiment 1, CD73 are stem cell surface marker of renal clear cell carcinoma
One, the acquisition of people's clear cell carcinoma of kidney lineage stem and functional verification
1, the acquisition of stem cell
1) get people's clear cell carcinoma of kidney clone 786-O cell, cultivate based on 5%CO with 1640 of the serum-free of interpolation 10% hyclone, 100IU/mL penicillin and 100g/mL streptomysin
237 DEG C of cultivations in incubator.
2) after step 1) being cultured to the 786-O cell trypsinization of exponential phase, resuspended with serum-free 1640 nutrient culture media containing 10ng/mL fibroblast growth factor, 20ng/mL epidermal growth factor, 5 μ g/mL insulin and 0.4% hyclone albumen again, cell suspension is entered 6 orifice plates without anchoring factor with 5000 cells/well kinds, within 7-10 days, forms micro-capsule.With after gentle cell separation liquid (Invitrogen, Eugene, OR, USA) re-suspended cell after enrichment micro-capsule cell, by normal 1640 medium culture containing serum, obtain the stem cell of 786-O.
2, the acquisition of ordinary cells
1) get people's clear cell carcinoma of kidney clone 786-O cell, cultivate based on 5%CO with containing 1640 of serum
237 DEG C of cultivations in incubator.
2) after step 1) being cultured to the 786-O cell trypsinization of exponential phase, more resuspended with 1640 nutrient culture media containing serum, enter to have 6 orifice plates of anchoring factor by cell suspension with 5000 cells/well kinds, carry out adhere-wall culture, obtain common 786-O cell.
3, the checking of 786-O stem cell
1) Serium-free Culture checking
By in step 1 2) after the stem cell trypsinization of 786-O that obtains, within 7-15 days, form the micro-capsule (as shown in Figure 1) of diameter about 200 μm afterwards by serum-free 1640 medium culture containing 10ng/mL fibroblast growth factor, 20ng/mL epidermal growth factor, 5 μ g/mL insulin and 0.4% hyclone albumen.
By in step 2 2) after the common 786-O cell trypsinization that obtains,, formed without micro-capsule after 7-15 days by serum-free 1640 medium culture containing 10ng/mL fibroblast growth factor, 20ng/mL epidermal growth factor, 5 μ g/mL insulin and 0.4% hyclone albumen.
To this demonstrate that in step 1 2) obtain the stem cell containing 786-O in micro-capsule cell.
2) NOD/SCID mouse tumor model method checking
Process: by step 1 2) after the stem cell trypsinization of 786-O that obtains, resuspended and adjust cell concentration to 5000 cells/mL with PBS, obtain cell suspension A; This cell suspension A is injected in 6-8 week ages female NOD/SCID mouse (Chinese Academy of Medical Sciences's Animal Experimental Study center) lower limb armpit on the left of, injection volume is 0.1mL(i.e. 500 cells).
Contrast: by step 2 2) after the common 786-O cell trypsinization that obtains, resuspended and adjust cell concentration to 5000 cells/mL with PBS, obtain cell suspension B; Be injected in by this cell suspension B on the right side of the same mouse lower limb armpit of above-mentioned process, injection volume is 0.1mL(i.e. 500 cells simultaneously).
Experiment establishes 3 repetitions, each repetition 5 mouse.
Result (as shown in Figure 2): inject after 120 days, by sacrifice, dissect mouse, the tumor formation rate of contrast is 0, and the tumor formation rate of process is 100%.
The result of experiment one shows, the micro-capsule cell that free serum culture is formed contains tumor stem cell.
Two, CD73 is the surface marker of clear cell carcinoma of kidney (786-O) stem cell
1, CD73 is the surface marker of clear cell carcinoma of kidney (786-O) stem cell
1) Western hybridization check
The common 786-O cell that the 786-O micro-capsule that in enrichment experiment one, step 1 obtains obtains with step 2 in experiment one, extract total protein of cell respectively and carry out Western hybridization, use CD73 monoclonal antibody (purchased from Abcam in hybridization, products catalogue is numbered ab59462) detect CD73 protein content, use β-actin monoclonal antibody (purchased from Abcam, products catalogue is numbered ab6276) detect β-actin protein content (contrast), as shown in Figure 3, in the stem cell of 786-O, CD73 protein content is apparently higher than common 786-O cell for result.
2) Immunofluorescence test
The 786-O micro-capsule that in enrichment experiment one, step 1 obtains, with containing the PBS of 0.5% hyclone soft wash twice after, with resuspended containing the PBS of 0.5% hyclone and adjust cell concentration to 1 × 10
5individual/mL, the fluorescently-labeled CD73 monoclonal antibody of PE is added (purchased from Abcam by the volume ratio of 400:1, products catalogue is numbered ab59462), ice bath washed twice with PBS after 30 minutes, the cell suspension obtained is dripped sheet, laser confocal microscope inspection is carried out with after DAPI mounting, result as shown in Figure 4, Fig. 4 from left to right the first width figure is the 786-O micro-capsule cell under bright field, second width figure is the result that under 340nm exciting light, 786-O micro-capsule cell core DAPI junction sends blue-fluorescence, 3rd width figure is the result that under 594nm exciting light, CD73 monoclonal antibody junction (i.e. CD73 location) sends red fluorescence, 4th width figure is the integrated results of the 3rd width and the 4th width figure, Fig. 4 result shows, CD73 is at 786-O micro-capsule cell surface high expressed.
3) Flow cytometry and sorting
Three repetitions are all established in following process, eachly repeat at least sorting 10000 cells.
Contrast: after 786-O stem cell trypsinization step 1 in experiment one obtained, uses the PBS containing 0.5% hyclone resuspended and adjusts cell concentration to 1 × 10
6individual/mL, after ice bath washed twice with the PBS of 0.5% hyclone after 30 minutes, is detected cell by BD FACSCalibur flow cytometer (manufacturer: BD company, model: Calibur).Result: fluorescence intensity is greater than 10
1.5cell proportion be 0.473%(such as shown in the A figure in Fig. 5).
Process 1: after 786-O stem cell trypsinization step 1 in experiment one obtained, uses the PBS containing 0.5% hyclone resuspended and adjusts cell concentration to 1 × 10
6individual/mL, the fluorescently-labeled CD73 monoclonal antibody of PE is added (purchased from Abcam by the volume ratio of 400:1, products catalogue is numbered ab59462), after ice bath washed twice with the PBS of 0.5% hyclone after 30 minutes, cell is detected by BD FACSCalibur flow cytometer (manufacturer: BD company, model: Calibur) and collects fluorescence intensity simultaneously respectively and be greater than 10
1.5cell (i.e. CD73 high expressing cell) and fluorescence intensity be less than 10
1.5cell (i.e. the low express cell of CD73).Result: fluorescence intensity is greater than 10
1.5cell proportion be 98.2%(such as shown in the B figure in Fig. 5).
Process 2: after 786-O ordinary cells trypsinization step 2 in experiment one obtained, uses the PBS containing 0.5% hyclone resuspended and adjusts cell concentration to 1 × 10
6individual/mL, the fluorescently-labeled CD73 monoclonal antibody of PE is added (purchased from Abcam by the volume ratio of 400:1, products catalogue is numbered ab59462), after ice bath washed twice with the PBS of 0.5% hyclone after 30 minutes, cell is detected by BD FACSCalibur flow cytometer (manufacturer: BD company, model: Calibur).Result: fluorescence intensity is greater than 10
1.5cell proportion be 0.331%(such as shown in the C figure in Fig. 5).
Step 1)-3) result show, CD73 can be used as the particular surface mark of 786-O stem cell.
2, the CD73 positive cell subgroup of sorting is 786-O stem cell
By in step 1 3) the CD73 high expressing cell of sorting is defined as CD73 positive cell, by step 1 3) the low expression of CD73 of sorting and not express cell be defined as CD73 negative cells.
1) NOD/SCID mouse tumor model method checking
By in step 1 3) the CD73 positive cell of separation and collection and negative cells adjust concentration with PBS respectively, obtains cell suspension X and X-that concentration is 5000 cells/mL respectively;
Get the female NOD/SCID mouse (Chinese Academy of Medical Sciences's Animal Experimental Study center) in 6-8 ages in week, on the right side of lower limb armpit, inject 0.1mL cell suspension X, on the left of lower limb armpit, inject 0.1mL cell suspension X-; Experiment establishes 3 repetitions, each repetition 5 mouse; Inject and put to death after 120 days, dissect mouse, result display CD73 negative cells tumor formation rate to be 0, CD73 positive cell tumor formation rate be 100%(as shown in Figure 6).
2) Serium-free Culture checking
By in step 1 3) the CD73 negative cells of separation and collection and positive cell be respectively with containing 10ng/mL fibroblast growth factor, 20ng/mL epidermal growth factor, serum-free 1640 medium culture of 5 μ g/mL insulin and 0.4% hyclone albumen, after 12 days, CD73 negative cells forms (as shown in the left figure in Fig. 7) without micro-capsule substantially, the micro-capsule (as shown in the right figure in Fig. 7) that CD73 positive cell is formed, statistics CD73 negative cells and positive cell form the ratio of micro-capsule, result as shown in Figure 8, it is 22% that the micro-capsule of CD73 positive cell forms average proportions, it is 2%(p=0.0007 that micro-capsule apparently higher than CD73 negative cells forms average proportions).
Result illustrates, CD73 positive cell is 786-O stem cell.
The expression of embodiment 2, CD73 is relevant to clear cell carcinoma of kidney grade malignancy (or clinical classification)
Clear cell carcinoma of kidney clinical sample source affiliated hospital of Lanzhou University second, all samples obtain after being clear cell carcinoma of kidney local or whole excision, and the acquisition of experiment sample meets the regulation of Gansu Ethics Committee.Wherein, normal kidney tissue sample size is 15, clear cell carcinoma of kidney sample size is 150.The sample acquisition time is 2010.9-2012.4.All sample population all do not accept radiation cure or chemotherapy.
1, clinical classification
First, the clear cell carcinoma of kidney grade malignancy (or clinical classification) of patient to be measured is divided into G1, G2 and G3 tri-grades according to following clinical criteria:
Described G1 level refers to that patient's clear cell carcinoma of kidney to be measured is limited in kidney peplos;
Described G2 level refers to that patient's clear cell carcinoma of kidney to be measured wears out kidney peplos, invades fat deposit, and is still confined in fascia renalis;
Described G3 level refers to that patient's clear cell carcinoma of kidney to be measured has invaded renal vein or/and inferior caval vein, invades adjacent organ or DISTANT METASTASES IN occurs.
According to above-mentioned standard, grade malignancy (or clinical classification) result recording each clinical sample is as shown in table 1.
2, CD73 positive cell ratio detects
Get a fritter nephridial tissue from clinical sample, fix, do histotomy after paraffin embedding with formalin, slice thickness is 4 μm.After section dimethylbenzene removes paraffin, ethanol purge, Steam Heating carries out antigen retrieval 20 minutes.Volume ratio and the CD73 monoclonal antibody (purchased from Abcam, products catalogue is numbered ab59462) of section being pressed 1:400 are hybridized, and diaminobenzidine dyes, haematoxylin redyeing.Statistics often to be opened in section CD73 positive cell number under random field and is accounted for the ratio of total cellular score in this visual field, and be that the results averaged ± standard deviation of G1, G2 and G3 lists in table 1 by grade malignancy, part coloration result as shown in Figure 9.Wherein, the coloration result of CD73 positive cell is nucleus is blue, and tenuigenin is brown; The coloration result of CD73 negative cells is nucleus is blue, and tenuigenin is colourless.Statistics tenuigenin is brown cell proportion, i.e. CD73 positive cell ratio.
The grade malignancy of table 1, sample and the positive ration statistics result of cell CD73
| Grade malignancy | Sample number | The ratio (%) of CD73 positive cell |
| G1 | 35 | 0.49±0.08 |
| G2 | 65 | 0.69±0.12 |
| G3 | 50 | 1.10±0.29 |
3, the standard of clear cell carcinoma of kidney grade malignancy is judged
According to the result of table 1, formulate and judge clear cell carcinoma of kidney grade malignancy (or clinical classification) standard or method as follows: when CD73 positive cell ratio in patient's nephridial tissue cell to be measured is 0.49% ± 0.08%(that is 0.41-0.57%) time, be described G1 level by described patient's clear cell carcinoma of kidney candidate to be measured; When CD73 positive cell ratio in patient's nephridial tissue cell to be measured is 0.69% ± 0.12%(that is 0.57%-0.81%) time, be described G2 level by described patient's clear cell carcinoma of kidney candidate to be measured; When CD73 positive cell ratio in patient's nephridial tissue cell to be measured is 1.10% ± 0.29%(that is 0.81%-1.39%) or higher than 1.39% time, be described G3 level by described patient's clear cell carcinoma of kidney candidate to be measured.
Embodiment 3, the expression of CD73 is utilized to judge clear cell carcinoma of kidney grade malignancy (or clinical classification)
1, the acquisition of sample to be tested
From the normal kidney tissue of Healthy People, sample size is 15; From the nephridial tissue of clear cell carcinoma of kidney patient, sample size is 25, and all samples obtain after being clear cell carcinoma of kidney local or whole excision, and the acquisition of experiment sample meets the regulation of Gansu Ethics Committee.All sample population all do not accept radiation cure or chemotherapy.
2, CD73 positive cell ratio detects
Carry out according to the method for step 2 in embodiment 2, in the nephridial tissue cell of 15 routine Healthy Peoples as a result, CD73 positive cell ratio is all extremely remarkable in 0.41%, and the result of the nephridial tissue of 25 routine clear cell carcinoma of kidney patients is as shown in table 2.
According to the standard of step 3 in embodiment 2, according to the clinical classification of CD73 positive cell ratio prediction clear cell carcinoma of kidney sample, outcome record is in " prediction grade malignancy " hurdle of table 2.
3, clinical classification
According to the clear cell carcinoma of kidney grade malignancy grade scale in step 1 in embodiment 2, the clear cell carcinoma of kidney grade malignancy of routine for 25 in step 1 patient is carried out classification, by outcome record in " actual measurement grade malignancy " hurdle of table 2.
The result of table 2. patient's clear cell carcinoma of kidney to be measured grade malignancy
The result of table 2 shows, by result according to the normative forecast clear cell carcinoma of kidney grade malignancy of embodiment 2 of the ratio that detects CD73 positive cell in nephridial tissue cell, has predicting the outcome of 23 examples to conform to measured result, rate of accuracy reached 92.0%.
In sum, CD73 can be used as the surface marker of clear cell carcinoma of kidney stem cell.And the clinical classification of the expression of CD73 and clear cell carcinoma of kidney is closely related, therefore, CD73 can be used for the molecular target of the diagnosis of the qualification of clear cell carcinoma of kidney stem cell and clear cell carcinoma of kidney, somatotype and detection, and can be used as the target gene of clear cell carcinoma of kidney biological therapy.
Claims (3)
1. detect the application of material in the product preparing diagnosis or auxiliary diagnosis patient's clear cell carcinoma of kidney to be measured grade malignancy of CD73.
2. application according to claim 1, is characterized in that: described grade malignancy is divided into G1, G2 and G3 tri-grades;
Described G1 level refers to that patient's clear cell carcinoma of kidney to be measured is limited in kidney peplos;
Described G2 level refers to that patient's clear cell carcinoma of kidney to be measured wears out kidney peplos, invades fat deposit, and is still confined in fascia renalis;
Described G3 level refers to that patient's clear cell carcinoma of kidney to be measured has invaded renal vein or/and inferior caval vein, invades adjacent organ or DISTANT METASTASES IN occurs.
3. application according to claim 2, is characterized in that:
The product of diagnosis or auxiliary diagnosis patient to be measured clear cell carcinoma of kidney grade malignancy comprises the carrier being described below diagnostic criteria: when in patient's nephridial tissue cell to be measured, CD73 positive cell ratio is 0.41-0.57%, be described G1 level by described patient's clear cell carcinoma of kidney candidate to be measured; When in patient's nephridial tissue cell to be measured, CD73 positive cell ratio is 0.57%-0.81%, be described G2 level by described patient's clear cell carcinoma of kidney candidate to be measured; When CD73 positive cell ratio in patient's nephridial tissue cell to be measured be 0.81%-1.39% or higher than 1.39% time, be described G3 level by described patient's clear cell carcinoma of kidney candidate to be measured.
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| CN111499747B (en) * | 2019-01-11 | 2022-03-18 | 康诺亚生物医药科技（成都）有限公司 | anti-CD 73 monoclonal antibody and application thereof |
| CN110018316B (en) * | 2019-04-16 | 2022-04-05 | 福建师范大学 | Application of DDX20 in preparation of renal clear cell carcinoma postoperative prognosis evaluation kit |
| CN116949168A (en) * | 2023-07-25 | 2023-10-27 | 河北医科大学第二医院 | Application of CD73 in diagnosis of parkinsonism |
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