Lentivirus-mediated short hairpin RNA interference of CENPK inhibits growth of colorectal cancer cells with overexpression of Cullin 4A

Table 1 Quality check results

Order number	Sample number	Sample name	Thermo NanoDrop 2000		2100 result		Results
			Concentration (ng/μL)	A260/A280	RIN	28S/18S
1	Y9148-1	NC	739.969	1.951	10	2.1	Pass
2	Y9148-2	NC	688.748	1.979	10	2.1	Pass
3	Y9148-3	NC	668.435	1.944	10	1.9	Pass
4	Y9149-1	KD	676.797	1.93	10	1.7	Pass
5	Y9149-2	KD	697.671	1.958	10	1.7	Pass
6	Y9149-3	KD	678.338	1.971	10	1.7	Pass

NC: RKO cells infected with centromere protein K negative control virus; KD: RKO cells with centromere protein K gene short hairpin RNA virus infection.

Table 2 Clustering of clinical data

Number	Variable	Group 1	Group 2
1	Age (yr)	< 57	> 57
2	Sex	Male	Female
3	T stage	T1/2/3	T4
4	N metastasis	N0	N1/2
5	Pathological grade	1/2	3
6	Clinical stage	I/II	III/IV

Table 3 Lentiviral-infected HCT 116 cells

Virus number	Experimental group	Virus titer	Virus infection dose
psc3741	NC (KD)	2.0E + 08 TU/mL	10.00 μL
LVpGCSIL-004PSC45249-1	KD	3.0E + 08 TU/mL	6.67 μL
KL9688-21	NC (OE)	4.0E + 8 TU/mL	5.00 μL
LVKL6733-2	OE	2.0E + 08 TU/mL	10.00 μL
LVpGCSIL-004PSC45249-1+ KL9688-21	KD+NC (OE)	3.0E + 08 + 2E + 8 TU/mL	3.33 + 5.00 μL
LVpGCSIL-004PSC45249-1+ LVKL6733-2	KD+OE	3.0E + 08 + 2E + 8 TU/mL	3.33 + 5.00 μL

NC: HCT 116 cells infected with centromere protein K negative control virus; KD: HCT 116 cells with centromere protein K gene short hairpin RNA virus infection; OE: HCT 116 cells virally infected with Cullin 4A.

Table 4 Expression and proportional statistics of the centromere protein K gene in cancerous and noncancerous tissue by immunohistochemical staining.

	Case group		P value
	Cancerous tissues	Noncancerous tissue
Higher expression	4	5	0.008
Lower expression	10	0

Table 5 Tissue type group * gene expression group using the Mann-Whitney U test

		CENPK gene expression group		Total	P value
		Low expression	High expression
Tissue type group	Cancerous tissues	146	160	306
	Noncancerous tissue	27	1	28	0.000
Total		173	161	334

CENPK: Centromere protein K.

Table 6 Clinical data * gene expression group analysis using the Mann-Whitney U test

		CENPK gene expression group		Total	P value
		Low expression	High expression
Sex	Male	88	103	191	0.460
	Female	58	57	115
Total		146	160	306
Age	< 57	75	82	157	0.983
	> 57	71	78	149
Total		146	160	306
T stage	T1/2/3	108	98	206	0.018
	T4	38	62	100
Total		146	160	306
N metastasis	N0	117	108	225	0.012
	N1/2	29	52	81
Total		146	160	306
Pathological grade	1/2	111	114	225	0.130
	3	29	45	74
Total		140	159	299
Clinical stage	I/II	116	108	224	0.019
	III/IV	30	52	82
Total		146	160	306

CENPK: Centromere protein K.

Table 7 Correlation between centromere protein K gene expression and clinical data in cancerous tissues

		Gene expression	T stage	N metastasis	Pathological grade
Gene expression	Correlation coefficient	1.000	0.135	0.143	0.135
	Significance (two-tailed)	0.000	0.018	0.012	0.018
T stage	Correlation coefficient	0.135	1.000	0.000	0.000
	Significance (two-tailed)	0.018	0.000	0.000	0.000
N metastasis	Correlation coefficient	0.143	0.000	1.000	0.000
	Significance (two-tailed)	0.012	0.000	0.000	0.000
Pathological grade	Correlation coefficient	0.135	0.000	0.000	1.000
	Significance (two-tailed)	0.018	0.000	0.000	0.000