Human papillomavirus type 16 E7 promotes cell viability and migration in cervical cancer by regulating the miR-23a/HOXC8 axis

Figures & data

Table 1. siRNA sequences used for cell transfection.

Name	Sequences (5′–3′)
Si-E6#1	CAGTTATGCACAGAGCTGCAAACAA
Si-E6#2	CACAGAGCTGCAAACAACTATACAT
Si-E7#1	CAACCAGAGACAACTGATCTCTACT
Si-E7#2	CAGAACCGGACAGAGCCCATTACAA
Si-E6/E7	GGUUGUGCGUACAAAGCAC
Si-NC	TTCTCCGAACGTGTCACGT

Table 2. Primer sequences used for qPCR.

Name	Sequences (5′–3′)
HPV16 E6 (forward)	AAGCAACAGTTACTGCGACGTGAGG
HPV16 E6 (reverse)	GGTCGGTGGACCGGTCGATGT
HPV16 E7 (forward)	CCAGCTGGACAAGCAGAACCGGA
HPV16 E7 (reverse)	GTGCCCCATCTGTTCTCAGAAACCA
HPV16 E6/E7 (forward)	CCGATTTCGGTTACGCCCTTAG
HPV16 E6/E7 (reverse)	CCATCTGTTCTCAGAAACC
HPV18 E6/E7 (forward)	AGCGCGCCATAGTATTGTGGT
HPV18 E6/E7 (reverse)	TTGCAGGAGGCAGCACAGAA
miR-23a (forward)	CGGATCACATTGCCAGGG
miR-23a (reverse)	CAGTGCGTGTCGTGGAGT
U6 (forward)	AAAGCAAATCATCGGACGACC
U6 (reverse)	GTACAACACATTGTTTCCTCGGA
HOXC8 (forward)	GAACCCGTGCTCGCTTAG
HOXC8 (reverse)	ACGCTCGCCTCTTGCTGA
GAPDH (forward)	GCAAGTTCAACGGCACAG
GAPDH (reverse)	CGCCAGTAGACTCCACGAC

Figure 1. E7 regulates the expression of miR-23a. (a) Expression of miR-23a in adjacent normal tissues, and HPV16+ and HPV16– tumour tissues. (b) Expression of miR-23a in CC cell lines (C33A, Hela, SiHa and CaSki). (c) Expression of E6/E7 in CC cell lines (C33A, Hela, SiHa and CaSki). (d) Transfection efficiency of E6 overexpression vector in C33A cells. (e) Transfection efficiency of E7 overexpression vector in C33A cells. (f) E6 mRNA and protein levels in SiHa and CaSki cells after si-E6#1 and si-E6#2 transfection. (g) E7 mRNA and protein levels in SiHa and CaSki cells after si-E7#1 and si-E7#2 transfection. (h) Expression of miR-23a in C33A cells after E6/E7, E6 and E7 overexpression. (i) Expression of miR-23a in SiHa and CaSki after E6/E7, E6 and E7 knockdown. **p < .01. ***p < .001.

Figure 2. E7 promotes the viability and migration of SiHa and CaSki cell lines by downregulating miR-23a expression. (a) The expression of miR-23a in SiHa and Hela cells after miR-23a mimic transfection. (b) Effects of miR-23a and E7 on CC cell viability were determined by CCK-8. Effects of miR-23a and E7 on CC cell migration were assessed using (c) Transwell assay and (d) wound healing assay. **p < .01 vs. the miR-nc + vector group. ^##p < .01 vs. the miR-23a + vector group.

Figure 3. miR-23a targets HOXC8. The targeting relationship (a) was predicted using the TargetScan database and (b) was confirmed by dual-luciferase reporter assay. (c) Effects of miR-23a and E7 on HOXC8 expression in SiHa and CaSki cells were detected using qPCR. (d) HOXC8 expression in normal and HPV16+ and HPV16– tumour tissues. (e) HOXC8 expression in CC cell lines (C33A, Hela, SiHa and CaSki). **p < .01. ^##p < .01 the miR-23a + vector group.

Data availability statement

All data included in this study are available upon request by contact with the corresponding author.