Use of protein depdc1 as a marker for diagnosing triple-negative breast cancer
A DEPDC1, triple-negative breast cancer technology, applied in the field of biomedicine, can solve the problems of poor diagnosis and treatment of triple-negative breast cancer, and achieve the effect of promoting colony formation, obvious technical effect, and positive technical effect
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Embodiment 1
[0045] Example 1 DEPDC1 is upregulated in three yin breast cancer
[0046] In order to clarify the role of DEPDC1 in three-yin breast cancer, the present invention analyzes the data of the two chips (GSE76250 and GSE81838) above the Gene Expression Omnibus (http: / / www.ncbi.nlm.nih.gov / geo), The expression of DEPDC1 in three yin breast cancer and pairing cancer is clarified. As figure 1 A and 1b show that DEPDC1 expression in tissue of Sanyin breast cancer is significantly increased compared to the paired cancer. Similar results can also be obtained in other databases.
[0047] In 33, the present invention has found that the expression of DEPDC1 in most three yin breast cancer tissues is higher than that of cancer. figure 1 C and 1D). Further, the expression of PCNA (nuclear proliferative antigen) in 165 cases of three-yin breast cancer tissues and Ki67 (a proliferative marker, the important indicator of tumor cell growth) is related to DEPDC1 expression (see figure 1 E and 1F). An...
Embodiment 2
[0048] Example 2 The DEPDC1 overduction in three yin breast cancer cells can promote cell proliferation and cloning.
[0049] The present invention then detected mRNA expression of DEPDC1 in a set of three-yin breast cancer cell lines. In these cells, the expression of endogenous DEPDC1 in MDA-MB-231 is significantly higher than that in other cell lines, while the expression of DEPDC1 in MDA-MB-436 cells is the lowest (see figure 2 A). Therefore, DEPDC1 was constructed in MDA-MB-436 cells to stabilize overexpress cell lines.
[0050] Real-time quantitative PCR and Western blot methods detect the mRNA and protein expression levels of DEPDC1 to confirm transfection efficiency ( figure 2 B and 2C). As image 3 A, the results of CCK-8 detection show that the cell activity increased significantly after DEPDC1. BRDU embedded experiments and PCNA expression were used to detect cell proliferation capabilities.
[0051] The present invention finds that DEPDC1 can increase the amount of BRDU...
Embodiment 3
[0054] Example 3 Three-yin breast cancer knockout DEPDC1 can reduce cell proliferation
[0055] In order to determine the physiological function of DEPDC1 in re-eremia, the present invention has also knocked down the expression of DEPDC in MDA-MB-231 cells. The efficiency of knockout is also determined by real-time quantitative PCR and Western blot ( Figure 4 A and Figure 4 B). like Figure 5 As shown in A, knocking DEPDC1 can inhibit the growth of MDA-MB-231 cells. Moreover, BRDU inhalation and PCNA protein expression levels were knocked out of DEPDC1 ( Figure 5B and 5C). The results of cell cycle distribution showed that the decrease in tumor cells in the Low DEPDC1 were increased (from 56.31% to 78.52%), and the S-phase cells were accompanied by 32.41% to 16.31%. Figure 5 D).
[0056] like Figure 5 E shows that the knockout DEPDC1 cloned in MDA-MB-231 cells was inhibited ( Figure 5 E). Moreover, the results of nude mice subcutaneous tumors show that knockout DEPDC1 significantly...
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