| Objective:The purpose of this study was to observe the expression level of FRα in CRC tissue,the level of FR+CTC in CRC patients and their relationship with clinical pathological factors,and explored whether FR+CTC had a role in the evaluation of the therapeutic efficacy of advanced CRC patients,providing novel biological molecular markers and new scientific measures for the early prevention and treatment of CRC and the clinical diagnosis and treatment monitoring of advanced CRC as well as further improving the prognosis of CRC.Method:The IHC was used to detect the expression of FRα in CRC and the corresponding adjacent tissues for comparative analysis.At the same time,peripheral blood of 33 CRC patients without anti-tumor therapy(defined as group A)and 8 patients with benign colorectal diseases(defined as group B)and 10 healthy people(defined as group C)were collected and tested for FR+CTC levels and a comparative analysis was also carried out.Correlation between initial FR+CTC and its clinicopathological factors in CRC patients was analyzed,the dynamic changes of FR+CTC numbers before and after chemotherapy in 15 patients with advanced CRC were observed,and the corresponding analysis of the therapeutic effect of drugs was carried out to verify the role and significance of FR+CTC in the evaluation of the efficacy of advanced CRC chemotherapy.The principle of the FRα positive CTC detection method is to use the CTC enrichment and purification system to remove red blood cells and most white blood cells in the sample after obtaining a 3m L patient blood sample and enrich the remaining rare cells in the sample including CTC.Then,specific small molecule probes were used to label and identify the highly expressed FRα on the surface of CTC.Finally,specific primers were used to bind Taqman probes,and polymerase chain reaction technology was used to quantitatively detect the small molecule probes bound by CTC,and the CTC content in the sample was evaluated by comparing the standard curves.Result:With IHC detection,the staining results showed that the positive rates of FRαexpression in 30 cases of CRC tissues and adjacent tissues were 87% and 37%,respectively,and the difference was statistically significant(P=0.0018).FRα-positive CTC detection method was used to detect the FR+CTC levels of 33 cases of CRC patients without anti-tumor therapy,namely group A,8 cases of benign colon disease,group B,and 10 healthy people,group C.Between group A and group B,the difference was statistically significant(P=0.0406).The difference between group A and group C was statistically significant(P<0.0001),while the difference between group B and group C was not statistically significant(P=0.2031).FR+CTC of the patient was analyzed by ROC curve diagnosis performance.With 8.5 FU/3 m L setting as the cutoff value,the area under the curve of FR+CTC diagnosis CRC was 0.8333(95% CI:0.7053-0.9613,P<0.0001),the sensitivity was 84.9% and the specificity is 88.9%.A total of 28 of 33 CRC patients were tested for CEA and CA19-9 at the same time.The sensitivity of FR+CTC detection was 84.8%,which was significantly higher than 53.6%of CA19-9 and 28.6% of CEA.Moreover,the baseline FR+CTC level of CRC patients in group A was not significantly related to age,gender,tumor location,T stage,and tumor differentiation(P>0.05),but was closely related to tumor N stage,M stage,and TNM stage.Patients with regional lymph node metastasis had higher FR+CTC levels than patients without regional lymph node metastasis,the difference was statistically significant(P=0.0048);M1 patients with distant metastasis had higher FR+CTC levels than those without distant metastasis Patients with M0 stage,and the difference was statistically significant(P=0.0145);and the FR+CTC level was related to the TNM stage.The FR+CTC level of patients with advanced CRC was higher than that of patients with early CRC(P=0.0217)The FR+CTC test kit was used to detect peripheral blood in 15 patients with advanced CRC before receiving chemotherapy or combined targeted drug therapy and during the corresponding first efficacy evaluation period.Among them,there were 11 patients with advanced CRC in first-line treatment,4 patients with advanced CRC in second-line treatment,and 4 patients with advanced CRC who received first-line treatment were evaluated as PD without clinical benefit and 7 patients with PR/SD patients had baseline FR+CTC levels(P=0.8970).However,after receiving treatment,the corresponding first efficacy evaluation was that the FR+CTC level of PD patients was increased from the baseline FR+CTC level,and the difference was statistically significant(P=0.0286).PR/SD patients FR+CTC level was significantly lower than the baseline FR+CTC level,and the difference was statistically significant(P=0.0379).The changes in FR+CTC levels before and after treatment were consistent with the results of treatment efficacy evaluation.Among patients with CRC who received second-line treatment,the efficacy evaluation was 2 patients with PD,and the level of FR+CTC increased from baseline after treatment,while the efficacy evaluation was 2 patients with SD patients,the level of FR+CTC decreased after treatment compared with baseline.And the change of FR+CTC level before and after treatment showed a consistent trend with the results of treatment efficacy evaluation.Conclusion:1.FRα should be a biological molecular marker for early diagnosis of CRC.2.It was suggested that FR+CTC should be used as a biological molecular marker for early diagnosis of CRC.FR+CTC might be used as a biological molecular marker to judge the malignant degree of CRC.3.FR+CTC might become a new measure for evaluating the efficacy of patients with advanced CRC receiving first-line drug therapy.FR+CTC also had potential application prospects in the efficacy evaluation of patients with advanced CRC receiving second-line drug therapy.However,it is worth further exploring that applying FR+CTC to CRC screening and becoming an evaluation measure for the efficacy of advanced CRC treatment still needs to be included in more cases for verification. |
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