CircLRP6调节miR-125a-5p/MCL1轴对食管鳞癌顺铂耐药性的影响

doi:10.3969/j.issn.1006-5725.2023.17.006

Abstract

Abstract:

Objective To investigate the impacts of circular RNA lipoprotein receptor 6 （circLRP6） on cisplatin （CDDP） resistance to esophageal squamous cell carcinoma （ESCC） and its relevant potential molecular mechanism. Methods CDDP?resistant cell lines of KYSE30 （KYSE30/CDDP?R） were established and divided into a control group， si?NC group， si?circLRP6 group， si?circLRP6 plus anti?NC group， and si?circLRP6 plus anti?miR?125a?5p group. The expression levels of circLRP6， miR?125a?5p and myeloid leukemia?1 （MCL?1） in tissues/cells were detected by RT?qPCR and Western blot. CCK?8 assay was applied to detect the CDDP sensitivity of KYSE30/CDDP?R cells， so was flow cytometry to detect apoptosis. Results mRNA levels of circLRP6 and MCL?1 were increased， while level of miR?125a?5p was decreased in ESCC cancer tissues/cells and CDDP?resistant cancer tissues （P < 0.001）. CircLRP6 silencing was able to up?regulate miR?125a?5p， inhibit the expression of MCL?1， enhance the sensitivity of CDDP?resistant ESCC cells to CDDP in vitro and in vivo， and promote the apoptosis of CDDP?resistant ESCC cells. Inhibition of miR?125a?5p weakened the effects of circLRP6 silencing on CDDP resistance and apoptosis in CDDP?resistant ESCC cells （P < 0.05）. circLRP6 could target and negatively regulate the expression of miR?125a?5p， and MCL?1 was the target of miR?125a?5p. Conclusion CircLRP6 silencing may enhance the sensitivity of CDDP?resistant ESCC cells to CDDP through the miR?125a?5p/MCL?1 axis.

Key words: circular RNA lipoprotein receptor 6, esophageal squamous cell carcinoma, cisplatin resistance, miR?125a?5p/MCL?1 axis

CLC Number:

R735.1

Pingping LIN,Fubo LI,Dongjuan WANG,Yan GUO,Yi. DONG. Impacts of circLRP6 on cisplatin resistance to esophageal squamous cell carcinoma by regulating miR⁃125a⁃5p/MCL1 axis[J]. The Journal of Practical Medicine, 2023, 39(17): 2183-2189.

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References 17

1	王静，杨雅阁，郑晓永，等. 安罗替尼单药在晚期食管鳞癌患者后线治疗中的疗效及安全性［J］. 实用医学杂志， 2022， 38（7）：893-899.
2	ZHOU T， FU H， DONG B， et al. HOXB7 mediates cisplatin resistance in esophageal squamous cell carcinoma through involvement of DNA damage repair［J］. Thorac Cancer， 2020， 11（11）：3071-3085.
3	YAJIMA S， SUZUKI T， NANAMI T， et al. Randomized phase II study to comparing docetaxel/nedaplatin versus docetaxel for 5-fluorouracil/cisplatin resistant esophageal squamous cell carcinoma［J］. Ann Thorac Cardiovasc Surg， 2021， 27（4）：219-224.
4	MU Q， LV Y， LUO C， et al. Research progress on the functions and mechanism of circRNA in cisplatin resistance in tumors［J］. Front Pharmacol， 2021， 12：709324.
5	WANG J， ZHU W， TAO G， et al. Circular RNA circ-LRP6 facilitates Myc-driven tumorigenesis in esophageal squamous cell cancer［J］. Bioengineered， 2020， 11（1）：932-938.
6	XU Y， ZHENG Y， DUAN Y， et al. MicroRNA-125a-5p targets LIM kinase 1 to inhibit cisplatin resistance of cervical cancer cells［J］. Oncol Lett， 2021， 21（5）：392.
7	LIU X， ZHAO T， YUAN Z， et al. MIR600HG sponges miR-125a-5p to regulate glycometabolism and cisplatin resistance of oral squamous cell carcinoma cells via mediating RNF44［J］. Cell Death Discov， 2022， 8（1）：216.
8	ZHAO Y， MA K， YANG S， et al. MicroRNA-125a-5p enhances the sensitivity of esophageal squamous cell carcinoma cells to cisplatin by suppressing the activation of the STAT3 signaling pathway［J］. Int J Oncol， 2018， 53（2）：644-658.
9	QI Y， DING Z， YAO Y， et al. Apigenin induces apoptosis and counteracts cisplatin-induced chemoresistance via Mcl-1 in ovarian cancer cells［J］. Exp Ther Med， 2020， 20（2）：1329-1336.
10	MICHELS J， OBRIST F， VITALE I， et al. MCL-1 dependency of cisplatin-resistant cancer cells［J］. Biochem Pharmacol， 2014， 92（1）：55-61.
11	YU X， LI W， XIA Z， et al. Targeting MCL-1 sensitizes human esophageal squamous cell carcinoma cells to cisplatin-induced apoptosis［J］. BMC Cancer， 2017， 17（1）：449-461.
12	JIA Y， TIAN C， WANG H， et al. Long non-coding RNA NORAD/miR-224-3p/MTDH axis contributes to CDDP resistance of esophageal squamous cell carcinoma by promoting nuclear accumulation of β-catenin［J］. Mol Cancer， 2021， 20（1）：162.
13	WANG Y， MO Y， PENG M， et al. The influence of circular RNAs on autophagy and disease progression［J］. Autophagy， 2022， 18（2）：240-253.
14	YU Y， DONG G， LI Z， et al. Circ-LRP6 contributes to osteosarcoma progression by regulating the miR-141-3p/HDAC4/HMGB1 axis［J］. Int J Oncol， 2022， 60（4）：38.
15	MISIR S， WU N， YANG B B. Specific expression and functions of circular RNAs［J］. Cell Death Differ， 2022， 29（3）：481-491.
16	CHANG N， GE N， ZHAO Y， et al. Hsa_circ_0007142 contributes to cisplatin resistance in esophageal squamous cell carcinoma via miR-494-3p/LASP1 axis［J］. J Clin Lab Anal， 2022， 36（5）：e24304.
17	TANG Y， YANG P， ZHU Y， et al. LncRNA TUG1 contributes to ESCC progression via regulating miR-148a-3p/MCL-1/Wnt/β- catenin axis in vitro［J］. Thorac Cancer， 2020， 11（1）：82-94.

RNA	Forward primer （5′-3′）	Reverse primer （5′-3′）
circLRP6	GAGTTGGATCAACCCAGAGC	TCCTCCAAGCCTCCAACTAC
LRP6	TTTAGCAAACAGACGGGACTT	GCCTCCAACTACAATCGTAGC
miR-125a-5p	CTGTCCCTGAGACCCTTTAAC	CGAGGAGAAGACGGAAGAAT
MCL-1	CGGCAGTCGCTGGAGATTAT	GTGGTGGTGGTTGGTTA
GAPDH	AGAAGGCTGGGGCTCATTTG	AGGGGCCATCCACAGTCTTC
U6	CTCGCTTCGGCAGCACA	ACGCTTCACGAATTTGCGTGTC

临床特征	例数	ESCC患者（n = 60）		P值
临床特征	例数	circLRP6高表达（n = 32）	circLRP6低表达（n = 28）	P值
性别
男性	35	21	14	0.336
女性	25	11	14
年龄
< 60岁	39	20	19	0.871
≥ 60岁	21	12	9
肿瘤大小
≤ 3 cm	32	19	13	0.457
> 3 cm	28	13	15
TNM分期
Ⅰ-Ⅱ	36	13	23	0.003
Ⅲ	24	19	5
淋巴结转移
无	20	6	14	0.022
有	40	26	14
CDDP敏感性
敏感的	33	11	22	0.002
耐药的	27	21	6

CDDP浓度（μmol/L）	细胞活力（%）
CDDP浓度（μmol/L）	KYSE30细胞	KYSE30/CDDP-R细胞
0	100.00 ± 0.00	100.00 ± 0.00
1	81.35 ± 4.57^*	97.72 ± 9.81
2	65.40 ± 4.10^*	91.58 ± 9.35
5	42.63 ± 3.95^*	78.30 ± 8.22^*
10	30.91 ± 3.42^*	65.45 ± 7.16^*
15	21.74 ± 2.83^*	51.20 ± 5.94^*
IC₅₀（μmol/L）	3.981	20.300

CDDP浓度（μmol/L）	细胞活力（%）
CDDP浓度（μmol/L）	Control	si-NC	si-circLRP6	si-circLRP6+anti-NC	si-circLRP6+anti-miR-125a-5p
0	100.00 ± 0.00	100.00 ± 0.00	100.00 ± 0.00	100.00 ± 0.00	100.00 ± 0.00
1	95.80 ± 9.64	96.20 ± 9.57	75.71 ± 8.05^*	79.99 ± 8.02^*	92.35 ± 9.18
2	88.75 ± 9.20	92.16 ± 9.14	61.43 ± 7.02^*	64.36 ± 6.15^*	81.64 ± 8.20^*
5	77.43 ± 8.15^*	80.25 ± 8.56^*	49.25 ± 6.01^*	48.28 ± 5.64^*	69.50 ± 6.93^*
10	64.92 ± 6.83^*	67.10 ± 7.08^*	41.96 ± 4.86^*	37.53 ± 5.01^*	54.71 ± 5.82^*
15	50.75 ± 5.42^*	51.33 ± 5.65^*	32.52 ± 3.74^*	30.41 ± 3.69^*	43.62 ± 5.01^*
IC₅₀（μmol/L）	20.114	21.325	4.982	4.854	12.150

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Impacts of circLRP6 on cisplatin resistance to esophageal squamous cell carcinoma by regulating miR⁃125a⁃5p/MCL1 axis

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