Low expression of Talin1 is associated with advanced pathological features in colorectal cancer patients

Abstract

To explore the proper prognostic markers for the likelihood of metastasis in CRC patients. Seventy-seven fresh CRC samples were collected to evaluate the mRNA level of the selected marker using Real-time PCR. Moreover, 648 formalin-fixed paraffin-embedded CRC tissues were gathered to evaluate protein expression by immunohistochemistry (IHC) on tissue microarrays. The results of Real-Time PCR showed that low expression of Talin1 was significantly associated with advanced TNM stage (p = 0.034) as well as gender (p = 0.029) in mRNA levels. Similarly, IHC results indicated that a low level of cytoplasmic expression of Talin1 was significantly associated with advanced TNM stage (p = 0.028) as well as gender (p = 0.009) in CRC patients. Moreover, decreased expression of cytoplasmic Talin1 protein was found to be a significant predictor of worse disease-specific survival (DSS) (p = 0.011) in the univariate analysis. In addition, a significant difference was achieved (p = 0.039) in 5-year survival rates of DSS: 65% for low, 72% for moderate, and 88% for high Talin1 protein expression. Observations showed that lower expression of Talin1 at both the gene and protein level may drive the disparity of CRC patients' outcomes via worse DSS and provide new insights into the development of progression indicators because of its correlation with increased tumor aggressiveness.

Figure 1

The common molecular markers between CTCs and exosome in databases. Fifteen hub genes were extracted from our mRNA and protein entrance mentioned databases.

Figure 2

KEGG pathway list related to the 15 excluded genes based on P-Value.

Figure 3

Talin1 plays significant roles in cancer related pathway.

Figure 4

The expression levels of Talin1 using Real-Time PCR in fresh tissue samples of CRC.

Figure 5

Box plot analysis of mRNA Talin1 expression levels in stage I to IV. Based on the standard definitions, each box-plot shows the median (bold line) and interquartile lines (box). The result of Mann–Whitney U test showed that there is an association for the median of Talin1 mRNA expression between stages I and III (p = 0.015) as well as stages II and III (p = 0.028). There were no statistically significant differences in the median level of Talin1 mRNA expression between other stages.

Figure 6

(A) Kaplan–Meier curves for disease-specific survival (DSS) and progression-free survival (PFS) based on mRNA Talin1 expression levels in CRC patients. In CRC patients, lower level of Talin1 mRNA expression was associated with shorter DSS compared to the tumors with high expression of Talin1 mRNA expression (P = 0.201). (B) Kaplan–Meier curve showed that low level of Talin1 expression is not significantly related to PFS (P = 0.698).

Figure 7

Immunohistochemical (IHC) analysis of Talin1 expression in different colorectal cancer (CRC) samples. CRC samples expressed Talin1 at various levels. Cytoplasmic expression of Talin1 at various levels: weak (A), moderate (B), and strong (C) in magnification of × 200; Weak (A-1), moderate (B-1), and strong (C-1) in magnification × 400. ECM expression of Talin1 in CRC at various levels is as follows: weak (D), moderate (E), and strong (F) in magnification × 200. Weak (D-1), moderate (E-1), and strong (F-1) in magnification × 400. IHC staining of kidney tissue was presented as positive (H) and negative (I) controls, adjacent normal tissue sample (G, G-1) in magnification × 200; and × 400.

Figure 8

Box plot analysis of Talin1 expression levels in stage I to IV. Based on the standard definitions, each box-plot shows the median (bold line) and interquartile lines (box). The result of Mann–Whitney U test showed that there is a statistically significant association for the median of cytoplasmic Talin1 expression between stages I and II (P = 0.013) as well as stages II and III (P = 0.001). There were no statistically significant differences in the median level of Talin1 protein expression between other stages.

Figure 9

Kaplan–Meier curves for disease-specific survival (DSS) and progression-free survival (PFS) based on cytoplasmic Talin1 protein expression level in CRC patients. (A) In CRC patients, lower level of Talin1 protein expression was associated with shorter DSS compared to the tumors with high expression of this protein (P = 0.048). (B) Kaplan–Meier survival analysis showed that low levels of Talin1 protein expression are not significantly related to PFS (P = 0.079).