Application of prothymosin alpha to preparation of breast cancer diagnosis marker

A technology for diagnosing markers and thymosin, which is applied in the field of tumor markers to achieve the effects of easy acquisition, easy popularization and application, and easy operation

Inactive Publication Date: 2014-11-12
XIAMEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Prothymosin alpha is a strongly acidic protein widely expressed in vivo, and its function is related to cellular immune regulation, cell proliferation and apoptosis [Wu C L, Shiau A L, Lin C C. Prothymosin alpha promotes cell proliferation in NIH3T3 cells[ J].Life Sci,1997,61:2091-2099], as a typical tumor-associated protein, in-depth research on the regulation of cell proliferation and apoptosis helps to discover the mechanism of cell carcinogenesis. It has been used in various cancers The increase of PTMA level was detected in [Zheng Jinlai, Liu Dianxin, Gao Lanxing. Research progress of prothymosin α[J]. Foreign Medical Immunology Volume, 1998, 21(4):210-214]. The PTMA protein product not only maintains cells in a highly proliferative s

Method used

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  • Application of prothymosin alpha to preparation of breast cancer diagnosis marker
  • Application of prothymosin alpha to preparation of breast cancer diagnosis marker
  • Application of prothymosin alpha to preparation of breast cancer diagnosis marker

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] Example 1 DOT-BLOT method was used to qualitatively detect the level of prothymosin alpha in the urine of tumor human and normal human.

[0019] Using the prepared prothymosin pro-alpha polyclonal antibody, the dot hybridization method was used to detect the level of pro-thymosin pro-alpha in the urine of tumor patients, and the urine of normal healthy people was used as the control (urine samples were all from Xiamen University Affiliated Zhongshan Hospital, provided by Liang Xianming, the technician in charge).

[0020] 1) Put 1 drop (2 μL) of urine on the NC membrane according to the table, and block it with blocking solution at 37°C for 2 hours after drying;

[0021] 2) Incubate with rabbit anti-thymosin pro-alpha (1:1000), overnight at 4°C;

[0022] 3) Wash the membrane with PBST for 3 times, shake on the shaker, 5min each time;

[0023] 4) HRP-labeled goat anti-rabbit secondary antibody (1:1000) was incubated at room temperature for 1 hour;

[0024] 5) Wash the...

Embodiment 2

[0026] Example 2 ELISA method was used to quantitatively detect the level of prothymosin alpha in the urine of tumor human and normal human.

[0027] Using the prepared prothymosin pro-alpha monoclonal antibody, using the indirect ELISA method, the urine in Example 1 was used as the detection object, and normal healthy human urine and PBS were used as controls to detect the level of pro-thymosin pro-alpha.

[0028] 1) Coating: The antigen was diluted 2X with Coating buffer according to Table 1, coated with 200 μL / well, protected from light, and kept overnight at 4°C.

[0029] 2) Wash the plate 3 times with PBST.

[0030] 3) Blocking buffer (5% skimmed milk powder) was added at 200 μL / Well, and blocked at 25° C. for 2 hours.

[0031] 4) Wash the plate 3 times with PBST, and spin dry the last time but do not buckle dry.

[0032] 5) Mouse anti-thymosin pro-alpha monoclonal antibody (1:1000) 200 μL / well, 2h at 25°C, protected from light.

[0033] 6) Wash the plate 3 times with ...

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Abstract

The invention relates to tumor markers, in particular to the application of prothymosin alpha to preparation of a breast cancer diagnosis marker. According to the application of prothymosin alpha to preparation of the breast cancer diagnosis marker, the dot blot hybridization method and the ELISA method are applied to detection of the level of prothymosin alpha protein in urine of a breast caner patient, and the relation between prothymosin alpha and the breast cancer biological behaviors is discussed; the immunoassay method is adopted, it is only required that the level of prothymosin alpha in urine is detected, it can quickly prove that the level of prothymosin alpha in urine of the breast cancer patient is obviously higher the normal level, preliminary judgment is conducted on breast cancer, and a basis is provided for further diagnosis; the needed urine sample is easy to obtain, any wound is avoided, an expensive instrument is not needed, operation is convenient, and popularization and application are easy.

Description

technical field [0001] The present invention relates to tumor markers, in particular to the application of prothymosin alpha in the preparation of breast cancer diagnostic markers. Background technique [0002] Breast cancer is one of the most common female malignancies, and the global incidence of breast cancer has been on the rise since the late 1970s [Yang L, Parkin DM, Ferlay J, et a1.Estimates of cancer incidence in China for 2000 and Projections for 2005 [J]. Cancer Epidemiol Biomarkers Prev, 2005, 14(1): 243-250]. One in eight women will suffer from breast cancer. China is not a country with a high incidence of breast cancer, but it should not be optimistic. This year, the growth rate of the incidence of breast cancer in my country is 1 to 2 percentage points higher than that of developed countries. According to the incidence data of breast cancer released by the National Cancer Center and the Bureau of Disease Control and Prevention of the Ministry of Health in 2012...

Claims

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Application Information

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IPC IPC(8): G01N33/574
CPCG01N33/57415G01N33/57484G01N2333/5759G01N2800/7028
Inventor 周克夫崔若辰梁贤明王世媛陈越月段莹陈艳芳张长弓陈雪飞孙雨涵胡鑫炎
Owner XIAMEN UNIV
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