Correlation of CDX2 with CD133， CD44 and its role in colorectal cancer

doi:10.3760/cma.j.cn371439-20260120-00105

Abstract

Abstract:

Objective To investigate the correlation， clinical relevance， and underlying molecular mechanisms associated with the expression of caudal type homeobox transcription factor 2 （CDX2） and the cancer stem cell （CSC） markers CD133 and CD44 in colorectal cancer （CRC）. Methods A total of 72 CRC patients who underwent surgical treatment at Luoyang Central Hospital from January 2016 to January 2018 were selected as the study subjects. The expression levels of CDX2， CD133， and CD44 in cancer tissues and normal tissues were compared； the clinical pathological characteristics and prognosis of patients with different CDX2， CD133， and CD44 expression were analyzed； univariate and multivariate Cox proportional hazards regression models were used to analyze prognostic factors； Pearson's chi-square test and Phi correlation analysis was used to test the correlation of expression between CDX2 and CD133， CD44. CDX2 in SW480 cells was silenced or overexpressed by lentivirus transfection； cell proliferation ability was detected by colony formation assay； migration ability was detected by scratch assay； invasion ability was detected by Transwell invasion assay； and the effect of CDX2 expression on CD133 and CD44 expression was analyzed by Western blotting. Results High expression of CDX2 accounted for 48.61% （35/72）， while low expression accounted for 51.39% （37/72）； both CD133 and CD44 showed high expression in 31.94% （23/72） of cases. There were statistically significant differences in T stage （χ²=6.18， P=0.013）， N stage （χ²=6.95， P=0.008）， and TNM stage （χ²=4.81， P=0.028） between patients with high and low CDX2 expression. In patients with high expression of both CD133 and CD44 compared to those with other expression patterns， there were also statistically significant differences in T stage （χ²=4.25， P=0.039）， N stage （χ²=5.91， P=0.015）， and TNM stage （χ²=5.13， P=0.024）. Multivariate analysis showed that low CDX2 expression was an independent risk factor simultaneously affecting recurrence free survival （RFS） and overall survival （OS） in CRC patients （RFS： HR=2.61， 95%CI： 1.18-5.77， P=0.018； OS： HR=2.69， 95%CI： 1.23-5.89， P=0.014）. Survival analysis showed that the 4-year RFS rates for CRC patients with low and high CDX2 expression were 32.07% and 85.53%， respectively， with a statistically significant difference （χ²=49.63， P<0.001）； the 4-year OS rates for CRC patients with low and high CDX2 expression were 21.62% and 82.86%， respectively， with a statistically significant difference （χ²=51.38， P<0.001）. The results of Pearson's chi-square test and Phi correlation analysis showed that the expression of CDX2 was significantly negatively correlated with CD133 （φ=-0.33， χ²=8.05， P=0.005）， CD44 （φ=-0.31， χ²=6.71， P=0.010） and CD133 combined with CD44 （φ=-0.49， χ²=17.12， P<0.001）. In CRC SW480 cells， the relative expression levels of CD133 in the blank control group， CDX2-silenced group， and CDX2-overexpressing group were 0.60±0.05， 0.93±0.05， and 0.42±0.06， respectively， and the relative expression levels of CD44 were 0.50±0.03， 0.77±0.08， and 0.35±0.03， respectively， with statistically significant differences （F=63.88， P<0.001； F=52.90， P<0.001）. Compared with the blank control group， the expression of CD133 and CD44 was significantly increased in the CDX2-silenced cells， while it was significantly decreased in the CDX2-overexpressing cells （all P<0.05）. Additionally， the expression levels of CD133 and CD44 in the CDX2-silenced cells were significantly higher than those in the CDX2-overexpressing cells （both P<0.05）. The numbers of cell clones in the blank control group， CDX2-silenced group， and CDX2-overexpressing group were 86.67±10.69， 206.00±17.52， and 44.33±6.51， respectively； the cell migration rates were 0.30±0.02， 0.51±0.03， and 0.22±0.03， respectively； and the numbers of invasive cells were 150.33±16.65， 312.67±28.29， and 65.67±12.22， respectively， with statistically significant differences （F=136.42， P<0.001； F=82.21， P<0.001； F=115.56， P<0.001）. Compared with the blank control group， the proliferation， migration， and invasion abilities of cells in the CDX2-silenced group were significantly enhanced （all P<0.05）， while those in the CDX2-overexpressing group were significantly reduced （all P<0.05）. The proliferation， migration， and invasion abilities of cells in the CDX2-silenced group were significantly stronger than those in the CDX2-overexpressing group （all P<0.05）. Conclusions Low CDX2 expression is an independent risk factor for poor prognosis in CRC， and CDX2 can serve as a prognostic marker for CRC. CDX2 expression is negatively correlated with the CSC markers CD133 and CD44. Silencing CDX2 significantly enhances the proliferative, migratory, and invasive capacities of SW480 cells, concomitant with upregulated expression of CD133 and CD44.

Key words: Colorectal neoplasms, Neoplastic stem cells, CDX2 transcription factor

Wang Yunshuai, Wang Huangjian, Wu Huibin, Zhang Hengyang, Xia Tianming. Correlation of CDX2 with CD133， CD44 and its role in colorectal cancer[J]. Journal of International Oncology, 2026, 53(11): 671-680.

Figures/Tables 14

References 24

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临床病理特征	CDX2高表达（n=35）	CDX2低表达（n=37）	χ²值	P值	CD133、CD44 双高表达（n=23）	非CD133、CD44 双高表达（n=49）	χ²值	P值
性别
男	19（54.29）	21（56.76）	0.04	0.833	11（47.83）	29（59.18）	0.82	0.366
女	16（45.71）	16（43.24）	0.04	0.833	12（52.17）	20（40.82）	0.82	0.366
年龄（岁）
≤60	21（60.00）	23（62.16）	0.04	0.851	17（73.91）	27（55.10）	2.33	0.127
>60	14（40.00）	14（37.84）	0.04	0.851	6（26.09）	22（44.90）	2.33	0.127
肿瘤部位
结肠	19（54.29）	20（54.05）	<0.01	0.984	11（47.83）	28（57.14）	0.55	0.459
直肠	16（45.71）	17（45.95）	<0.01	0.984	12（52.17）	21（42.86）	0.55	0.459
T分期
T_1-2	15（42.86）	6（16.22）	6.18	0.013	3（13.04）	18（36.73）	4.25	0.039
T_3-4	20（57.14）	31（83.78）	6.18	0.013	20（86.96）	31（63.27）	4.25	0.039
N分期
N₀	25（71.43）	15（40.54）	6.95	0.008	8（34.78）	32（65.31）	5.91	0.015
N_1-2	10（28.57）	22（59.46）	6.95	0.008	15（65.22）	17（34.69）	5.91	0.015
M分期
M₀	34（97.14）	33（89.19）	0.75	0.388	19（83.61）	48（97.96）	3.58	0.059
M₁	1（2.86）	4（10.81）	0.75	0.388	4（17.39）	1（2.04）	3.58	0.059
TNM分期
Ⅰ~Ⅱ	25（71.43）	17（45.95）	4.81	0.028	9（39.13）	33（67.35）	5.13	0.024
Ⅲ~Ⅳ	10（28.57）	20（54.05）	4.81	0.028	14（60.87）	16（32.65）	5.13	0.024
肿瘤大小（cm）
<3	22（62.86）	15（40.54）	3.59	0.058	12（52.17）	25（51.02）	0.01	0.927
≥3	13（37.14）	22（59.46）	3.59	0.058	11（47.83）	24（48.98）	0.01	0.927
分化程度
高-中分化	32（91.43）	32（86.49）	0.09	0.770	22（95.65）	42（85.71）	0.72	0.396
低-未分化	3（8.57）	5（13.51）	0.09	0.770	1（4.35）	7（14.29）	0.72	0.396

变量	单因素分析		多因素分析
变量	HR值（95%CI）	P值	HR值（95%CI）	P值
性别（男/女）	1.45（0.76~2.77）	0.265
年龄（≤60岁/>60岁）	0.68（0.35~1.32）	0.228
T分期（T_1-2/T_3-4）	0.35（0.18~0.69）	0.013	0.68（0.24~1.99）	0.485
N分期（N₀/N_1-2）	0.40（0.20~0.79）	0.004	0.64（0.13~3.25）	0.587
TNM分期（Ⅰ~Ⅱ/Ⅲ~Ⅳ）	0.45（0.23~0.88）	0.011	0.85（0.16~4.60）	0.850
肿瘤部位（结肠/直肠）	1.40（0.73~2.66）	0.309
肿瘤大小（<3 cm/≥3 cm）	0.54（0.28~1.04）	0.055
分化程度（高-中分化/低-未分化）	0.12（0.03~0.49）	0.003	0.23（0.08~0.63）	0.004
CDX2（低表达/高表达）	4.07（2.09~7.95）	<0.001	2.61（1.18~5.77）	0.018
CD133（低表达/高表达）	0.49（0.26~0.94）	0.042	0.49（0.21~1.12）	0.091
CD44（低表达/高表达）	0.42（0.22~0.80）	0.012	0.45（0.20~1.01）	0.053

变量	单因素分析		多因素分析
变量	HR值（95%CI）	P值	HR值（95%CI）	P值
年龄（≤60岁/>60岁）	0.72（0.38~1.36）	0.288
性别（男/女）	1.35（0.73~2.49）	0.342
肿瘤部位（结肠/直肠）	1.15（0.62~2.13）	0.648
肿瘤大小（<3 cm/≥3 cm）	0.63（0.34~1.16）	0.125
分化程度（高-中分化/低-未分化）	0.19（0.05~0.72）	0.014	0.32（0.12~0.87）	0.025
T分期（T_1-2/T_3-4）	0.39（0.20~0.73）	0.015	0.53（0.17~1.65）	0.273
N分期（N₀/N_1-2）	0.33（0.18~0.64）	<0.001	0.25（0.05~1.19）	0.082
TNM分期（Ⅰ~Ⅱ/Ⅲ~Ⅳ）	0.45（0.23~0.85）	0.007	0.47（0.09~2.39）	0.364
CDX2（低表达/高表达）	4.48（2.38~8.41）	<0.001	2.69（1.23~5.89）	0.014
CD133（低表达/高表达）	0.44（0.24~0.82）	0.016	0.55（0.25~1.22）	0.142
CD44（低表达/高表达）	0.50（0.27~0.93）	0.037	0.60（0.29~1.24）	0.165

组别	CD133	CD44
空载对照组	0.60±0.05	0.50±0.03
CDX2沉默组	0.93±0.05^a	0.77±0.08^a
CDX2过表达组	0.42±0.06^ab	0.35±0.03^ab
F值	63.88	52.90
P值	<0.001	<0.001

组别	细胞克隆形成数（个）	细胞迁移率	侵袭细胞数（个）
空载对照组	86.67±10.69	0.30±0.02	150.33±16.65
CDX2沉默组	206.00±17.52^a	0.51±0.03^a	312.67±28.29^a
CDX2过表达组	44.33±6.51^ab	0.22±0.03^ab	65.67±12.22^ab
F值	136.42	82.21	115.56
P值	<0.001	<0.001	<0.001