Serum hsa_circ_0087776 as a new oncologic marker for the joint diagnosis of multiple myeloma

Figures & data

Figure 1. Methodological evaluation of hsa _circ _0087776. (a) The hsa_circ_0087776 standard curve. (b) The 18s standard curve. (c, d) Stability of hsa_circ_0087776 and 18s in room temperature incubation and repeated freeze-thaw

Table 1. The intra-assay and interassay repeatability difference of hsa_circ_0087776

	hsa _circ _0087776	18s
Intra-assay
Mean ± SD	29.797 ± 0.358	19.650 ± 0.278
CV, %	1.07%	0.83%
Inter-assay
Mean ± SD	30.209 ± 0.5576	17.823 ± 0.353
CV, %	1.67%	1.06%

Note: The intra-assay and inter-assay of hsa_circ_0087776 and 18 s were lower than 2%. CV: Coefficient of Variance.

Figure 2. Methodological evaluation of hsa _circ _0087776. (a) Single peak specificity of hsa _circ _0087776 and 18s dissolution and amplification curves. (b) The specificity of hsa _circ _0087776 and 18s was verified by agarose gel electrophoresis. (c) Sequencing results of hsa _circ _0087776 and 18s

Figure 3. Detection of hsa _circ _0087776 relative expression in serum. (a) Hsa_circ-0087776 and 18s mRNA expression after treatment with RNase R. (b) Differential expression of hsa-circ-0087776 in serum of MM patients. (c) Differential expression of hsa-circ-0087776 in MM cell lines. (*P < 0.05; **P < 0.01; ***P < 0.001)

Figure 4. Evaluation of combined diagnostic efficacy of hsa _circ _0087776. (a) The ROC curve of serum hsa _circ _0087776. (b) The ROC curve of ALB. (c) The ROC curve of β2-MG. (d) By contrast, the diagnostic efficacy of serum hsa _circ _0087776 is remarkable

Figure 5. Changes in the expression of hsa_circ_0087776 before and after treatment (P = 0.0111)

Table 2. Use of Hsa_circ_ 0087776, ALB, and β₂-MG levels to distinguish MM patients from healthy participants

	SEN (%)	SPE (%)	ACCU (%)	PPV (%)	NPV (%)
β₂-MG	32.4%	83.8%	50.5%	78.6%	40.3%
	(44/136)	(62/74)	(106/210)	(44/56)	(62/154)
Alb	33.1%	47.3%	38.1%	53.6%	27.8%
	(45/136)	(35/74)	(80/210)	(45/84)	(35/126)
Hsa_circ_ 0087776	72.8%	67.6%	71.0%	80.5%	57.5%
	(99/136)	(50/74)	(149/210)	(99/123)	(50/87)
β₂-MG + Hsa_circ_ 0087776	82.4%	56.8%	73.3%	77.8%	63.6%
	(112/136)	(42/74)	(154/210)	(112/144)	(42/66)
Alb + Hsa_circ_ 0087776	83.1%	35.1%	66.2%	70.0%	53.1%
	(113/136)	(26/74)	(139/210)	(113/162)	(26/49)
Alb + Hsa_circ_ 0087776 + β₂-MG	88.2%	31.1%	68.1%	70.2%	41.0%
	(120/136)	(23/74)	(143/210)	(120/171)	(16/39)

Note: Sensitivity (SEN); specificity (SPE); overall accuracy (ACCU); positive predictive value (PPV); negative predictive value (NPC)

Table 3. Correlation between hsa_circ_ 0087776 expression and clinicopathologic features of MM patients

Clinical Characteristics	Case Number	hsa_circ_ 0087776		Pearson χ^2	P-value
		Low Expression	High Expression
Gender				2.944	0.860
Male	66	38	28
Female	70	30	40
Age				0.268	0.605
<65	75	39	36
≥65	61	29	32
M protein				1.121	0.290
IgA	84	39	45
IgG	52	29	23
Light chain				0.472	0.492
λ	64	30	34
κ	72	38	34
HGB (g/L)				0.138	0.710
<120	94	46	48
≥120	42	22	20
Total protein				0.287	0.592
Normal	97	42	45
Abnormal	49	26	23
Albumin				0.538	0.463
Normal	92	44	48
Abnormal	44	24	20
Globulin				0.282	0.595
Normal	85	41	44
Abnormal	51	27	24
β₂-MG (μg/L)				0.034	0.854
<3.5	93	46	47
≥3.5	43	22	21
Renal injury				4.225	0.040*
Yes	41	26	15
No	95	42	53
Bone injury				0.033	0.857
Yes	89	45	44
No	47	23	24

Note: Statistical analyses were carried out using Pearson χ² test. *P < 0.05 was considered significant

Table 4. Univariate and multivariate analysis of MM-ISS staging and prognostic factors

Clinicopathological Parameters	Univariate analysis		Multivariate analysis
	HR (95% CI)	P-value	HR (95% CI)	P-value
β₂-MG (<3.5 μg/ml vs ≥3.5 μg/ml)	2.471 (1.403, 4.351)	0.002**	2.711 (1.394, 5.271)	0.003**
Albumin (Outlier interval vs Rrference interval)	0.429 (0.265, 0.696)	0.001**	0.327 (0.183, 0.585)	< 0.001***
Renal injury (Yes vs No)	1.088 (0.865, 1.396)	0.477	-	-
Bone injury (Yes vs No)	5.482 (3.289, 9.136)	< 0.001***	5.224 (2.971, 9.187)	< 0.001***
hsa_circ_0087776 (low expresssion vs high expression)	4.228 (2.564, 6.974)	< 0.001***	4.191 (2.371, 7.406)	< 0.001***

Note: Univariate and multivariate analyses were performed by logistic regression analysis.

*P < 0.05, **P < 0.01, ***P < 0.001. CI:confidence interval.

Data availability statement

The datasets used and/or analyzed during the current study are available from the corresponding author on reasonable request.